Indole AHR inhibitors and uses thereof

ABSTRACT

The present invention provides compounds useful as inhibitors of AHR, compositions thereof, and methods of using the same.

BACKGROUND OF THE INVENTION

The aryl hydrocarbon receptor (AHR) is a transcription factor thatwithout ligand exists in the inactive state in the cytoplasm bound toHSP90. Upon ligand binding, AHR translocates to the nucleus where itdimerizes with ARNT forming a functional transcription factor. AHR/ARNTbinds dioxin response elements (DRE) in the promotor of many genes whereit modulates gene transcription. The most well documented genesregulated by AHR are the cytochrome P450 genes Cyp1b1 and Cyp1a1, whereactivation of AHR greatly increases expression of these genes.Therefore, Cyp1b1 and Cyp1a1 mRNA levels are a selective readout of AHRactivation (reviewed in Murray et al., 2014).

Many exogenous and endogenous agonists of AHR exist that activate thereceptor. The best characterized exogenous ligand class are the dioxins.One of the first endogenous ligands to be characterized is kynurenine,generated by TDO (Opitz 2011) or IDO (Mezrich 2010). Kynurenine is astable metabolite in the IDO/TDO pathway and is the product oftryptophan degradation. Kynurenine has been shown to activate AHR asmeasured by an increase in Cyp1a1 and/or Cyp1b1 mRNA levels in multiplecell types, along with other DRE-driven genes.

AHR activation has pro-tumor effects by acting directly on the tumorcells and indirectly by causing immunosuppression, therefore notallowing the body's own immune system to attack the tumor. For example,AHR activation through multiple ligands leads to increased expression ofFoxP3 and results in a polarization of CD4+ T-cells toward a suppressivesubset called Foxp3+ T-regulatory cells (Tregs). These T-reg cellsinhibit the proliferation of activated T cells (Funatake 2005, otherrefs). Interestingly, kynurenine has been shown to induceimmunosuppressive Tregs through AHR. Kynurenine does not affect T-reggeneration in AHR-null T cells or when an AHR antagonist is added(Mezrich). In addition to T-regs, AHR activation also leads to expansionof suppressive Tr1 T cells (Gandhi 2010). It has also been shown thatexpression of IDO is regulated by AHR activation in both tumor cells andT cells, leading to increased immune suppression (Vogel). It is likelythere is also a role for AHR in immune suppressive myeloid cells (Nguyen2013). Immune suppression is often associated with high levels ofanti-inflammatory cytokines and there is evidence that AHR is involvedin activation of many of these cytokines, such as IL-10 (Gandhi 2010,Wagage 2014).

There remains an unmet need to develop inhibitors of AHR for treatingdiseases, disorders and conditions associated therewith.

SUMMARY OF THE INVENTION

It has now been found that compounds of this invention, andpharmaceutically acceptable compositions thereof, are effective asinhibitors of AHR. Such compounds have the general formula I:

or a pharmaceutically acceptable salt thereof, wherein each variable isas defined and described herein.

Compounds of the present invention, and pharmaceutically acceptablecompositions thereof, are useful for treating a variety of diseases,disorders or conditions, associated with AHR. Such diseases, disorders,or conditions include those described herein.

Compounds provided by this invention are also useful for the study ofAHR in biological and pathological phenomena; the study of intracellularsignal transduction pathways; and the comparative evaluation of new AHRinhibitors in vitro or in vivo.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts a plot showing inhibition of IL22 and cyp1a1 RNAexpression levels with a compound described herein.

FIG. 2 depicts a plot showing decrease in IL-22 protein (top) andincrease in IL-2 (bottom) by treatment of activated T cells with acompound described herein.

FIG. 3 depicts a plot showing CT26 efficacy study with a vehicle, acompound described herein, anti-PD-1 and a combination thereof.

FIG. 4 depicts a plot showing B16-IDO efficacy study with a vehicle, acompound described herein, anti-PD-1 and a combination thereof.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS 1. General Description ofCompounds of the Invention

In certain embodiments, the present invention provides inhibitors ofAHR. In some embodiments, such compounds include those of formula I:

or a pharmaceutically acceptable salt thereof, wherein:Ring A is selected from:

-   each R¹ is independently selected from R, —C(O)R, —C(O)OR, —SO₂R,    —C(O)N(R)₂, or —SO₂RN(R)₂;-   each R is independently hydrogen, deuterium, or an optionally    substituted group selected from C₁₋₆ aliphatic, a 3-8 membered    saturated or partially unsaturated monocyclic carbocyclic ring,    phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring; a 4-8    membered saturated or partially unsaturated monocyclic heterocyclic    ring having 1-2 heteroatoms independently selected from nitrogen,    oxygen, or sulfur, a 5-6 membered monocyclic heteroaromatic ring    having 1-4 heteroatoms independently selected from nitrogen, oxygen,    or sulfur, or an 8-10 membered bicyclic heteroaromatic ring having    1-5 heteroatoms independently selected from nitrogen, oxygen, or    sulfur;-   each or R^(x), R^(y), and R^(z) is independently selected from R,    halogen, cyano, nitro, —OR, —SR, —N(R)₂, —N(R)C(O)R, —C(O)N(R)₂,    —N(R)C(O)N(R)₂, —N(R)C(O)OR, —OC(O)N(R)₂, —N(R)SO₂R, —SO₂RN(R)₂,    —C(O)R, —C(O)OR, —OC(O)R, —C(O)OR, —S(O)R, or —SO₂R, or:    -   two R^(x) on the same carbon are taken together to form ═O or        ═S; or:    -   two R^(y) on the same carbon are taken together to form ═O or        ═S;-   each of m and n is independently 1, 2, 3, 4, or 5;-   Ring B is phenyl, a 5-6 membered monocyclic heteroaromatic ring    having 1-4 heteroatoms independently selected from nitrogen, oxygen,    or sulfur, or an 8-10 membered bicyclic heteroaryl ring having 1-4    heteroatoms independently selected from nitrogen, oxygen, or sulfur;-   Ring C is phenyl or a 5-6 membered heteroaryl ring having 1-4    heteroatoms independently selected from nitrogen, oxygen, or sulfur;    or an 8-10 membered bicyclic heteroaryl ring having 1-4 heteroatoms    independently selected from nitrogen, oxygen, or sulfur;-   L¹ is a covalent bond or an optionally substituted C₁₋₆ membered    straight or branched bivalent hydrocarbon chain wherein a methylene    unit of L¹ is optionally replaced with -Cy-, —O—, —S—, —NR—, —C(O)—,    —C(O)O—, —OC(O)—, —C(O)N(R)—, —N(R)C(O)—, —SO₂—, —N(R)SO₂—, or    SO₂N(R)—S; and-   -Cy- is a 3-8 membered bivalent saturated, partially unsaturated, or    aromatic monocyclic ring having 0-2 heteroatoms independently    selected from nitrogen, oxygen, or sulfur, or an 8-10 membered    bivalent saturated, partially unsaturated, or aromatic bicyclic ring    having 0-3 heteroatoms independently selected from nitrogen, oxygen,    or sulfur.

In some embodiments, the present invention provides inhibitors of AHR,such compounds include those of formula I′:

or a pharmaceutically acceptable salt thereof, wherein:

-   Ring A is selected from:

-   each p is independently 0, 1, or 2, as valency will allow;-   each R¹ is independently selected from R, —C(O)R, —C(O)OR, —SO₂R,    —C(O)N(R)₂, or —SO₂RN(R)₂;-   each R is independently hydrogen, deuterium, or an optionally    substituted group selected from C₁₋₆ aliphatic, a 3-8 membered    saturated or partially unsaturated monocyclic carbocyclic ring,    phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring; a 4-8    membered saturated or partially unsaturated monocyclic heterocyclic    ring having 1-2 heteroatoms independently selected from nitrogen,    oxygen, or sulfur, a 5-6 membered monocyclic heteroaromatic ring    having 1-4 heteroatoms independently selected from nitrogen, oxygen,    or sulfur, or an 8-10 membered bicyclic heteroaromatic ring having    1-5 heteroatoms independently selected from nitrogen, oxygen, or    sulfur; or two R on the same nitrogen are taken together with their    intervening atoms to form a 4-7 membered saturated, partially    unsaturated, or aromatic ring having 1-2 heteroatoms in addition to    the nitrogen independently selected from oxygen, nitrogen, or    sulfur;-   each or R^(x), R^(y), and R^(z) is independently selected from R,    halogen, cyano, nitro, —OR, —SR, —N(R)₂, —N(R)C(O)R, —C(O)N(R)₂,    —C(O)N(R)OR, —N(R)C(O)N(R)₂, —N(R)C(O)OR, —OC(O)N(R)₂, —N(R)SO₂R,    —SO₂RN(R)₂, —C(O)R, —C(O)OR, —OC(O)R, —C(O)OR, —S(O)R, or —SO₂R, or:-   two R^(x) on the same carbon are taken together to form ═O or ═S;    or:-   two R^(y) on the same carbon are taken together to form ═O or ═S;-   each of m and n is independently 1, 2, 3, 4, or 5;-   Ring B is phenyl, a 7-10 membered bicyclic partially unsaturated or    aromatic carbocyclic ring, a 5-6 membered monocyclic heteroaromatic    ring having 1-4 heteroatoms independently selected from nitrogen,    oxygen, or sulfur, an 8-10 membered bicyclic heteroaryl ring having    1-4 heteroatoms independently selected from nitrogen, oxygen, or    sulfur, or a 12-15 membered partially unsaturated or aromatic    tricyclic ring having 1-4 heteroatoms independently selected from    nitrogen, oxygen, or sulfur;-   Ring C is phenyl or a 5-6 membered heteroaryl ring having 1-4    heteroatoms independently selected from nitrogen, oxygen, or sulfur;-   L¹ is a covalent bond or an optionally substituted C₁₋₆ membered    straight or branched bivalent hydrocarbon chain wherein a methylene    unit of L¹ is optionally replaced with -Cy-, —O—, —S—, —NR—, —C(O)—,    —C(O)O—, —OC(O)—, —C(O)N(R)—, —N(R)C(O)—, —SO₂—, —N(R)SO₂—, or    SO₂N(R)—S; and-   Cy- is a 3-8 membered bivalent saturated, partially unsaturated, or    aromatic monocyclic ring having 0-2 heteroatoms independently    selected from nitrogen, oxygen, or sulfur, or an 8-10 membered    bivalent saturated, partially unsaturated, or aromatic bicyclic ring    having 0-3 heteroatoms independently selected from nitrogen, oxygen,    or sulfur.

In some embodiments, the present invention provides a compound offormula I or formula I′, with the proviso that when Ring A is

Ring B is not

and/or Ring C is not

and/or R¹ is not

As generally defined above, Ring A is selected from:

One of ordinary skill in the art would readily understand and appreciatethat there are multiple orientations of Ring A. For example, and for thepurposes of clarity, when Ring A is selected to be

embodiments may be envisioned whereby Ring A is oriented in formula I orformula I′ as

Accordingly, both such orientations are contemplated by the presentinvention.

In some embodiments, the present invention provides a compound offormula I or formula I′, with the proviso that L¹ is not —NHCH₂CH₂—. Insome embodiments, the present invention provides a compound of formula Ior formula I′, with the proviso that when Ring A is

L¹ is not —NHCH₂CH₂—.

2. Compounds and Definitions

Compounds of this invention include those described generally above, andare further illustrated by the classes, subclasses, and speciesdisclosed herein. As used herein, the following definitions shall applyunless otherwise indicated. For purposes of this invention, the chemicalelements are identified in accordance with the Periodic Table of theElements, CAS version, Handbook of Chemistry and Physics, 75^(th) Ed.Additionally, general principles of organic chemistry are described in“Organic Chemistry”, Thomas Sorrell, University Science Books,Sausalito: 1999, and “March's Advanced Organic Chemistry”, 5^(th) Ed.,Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York: 2001, theentire contents of which are hereby incorporated by reference.

The term “aliphatic” or “aliphatic group”, as used herein, means astraight-chain (i.e., unbranched) or branched, substituted orunsubstituted hydrocarbon chain that is completely saturated or thatcontains one or more units of unsaturation, or a monocyclic hydrocarbonor bicyclic hydrocarbon that is completely saturated or that containsone or more units of unsaturation, but which is not aromatic (alsoreferred to herein as “carbocycle,” “cycloaliphatic” or “cycloalkyl”),that has a single point of attachment to the rest of the molecule.Unless otherwise specified, aliphatic groups contain 1-6 aliphaticcarbon atoms. In some embodiments, aliphatic groups contain 1-5aliphatic carbon atoms. In other embodiments, aliphatic groups contain1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groupscontain 1-3 aliphatic carbon atoms, and in yet other embodiments,aliphatic groups contain 1-2 aliphatic carbon atoms. In someembodiments, “cycloaliphatic” (or “carbocycle” or “cycloalkyl”) refersto a monocyclic C₃-C₆ hydrocarbon that is completely saturated or thatcontains one or more units of unsaturation, but which is not aromatic,that has a single point of attachment to the rest of the molecule.Suitable aliphatic groups include, but are not limited to, linear orbranched, substituted or unsubstituted alkyl, alkenyl, alkynyl groupsand hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or(cycloalkyl)alkenyl.

The term “lower alkyl” refers to a C₁₋₄ straight or branched alkylgroup. Exemplary lower alkyl groups are methyl, ethyl, propyl,isopropyl, butyl, isobutyl, and tert-butyl.

The term “lower haloalkyl” refers to a C₁₋₄ straight or branched alkylgroup that is substituted with one or more halogen atoms.

The term “heteroatom” means one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon (including, any oxidized form of nitrogen,sulfur, phosphorus, or silicon; the quaternized form of any basicnitrogen or; a substitutable nitrogen of a heterocyclic ring, forexample N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) orNR⁺ (as in N-substituted pyrrolidinyl)).

The term “unsaturated,” as used herein, means that a moiety has one ormore units of unsaturation.

As used herein, the term “bivalent C₁₋₈ (or C₁₋₆) saturated orunsaturated, straight or branched, hydrocarbon chain”, refers tobivalent alkylene, alkenylene, and alkynylene chains that are straightor branched as defined herein.

The term “alkylene” refers to a bivalent alkyl group. An “alkylenechain” is a polymethylene group, i.e., —(CH₂)_(n)—, wherein n is apositive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from1 to 2, or from 2 to 3. A substituted alkylene chain is a polymethylenegroup in which one or more methylene hydrogen atoms are replaced with asubstituent. Suitable substituents include those described below for asubstituted aliphatic group.

The term “alkenylene” refers to a bivalent alkenyl group. A substitutedalkenylene chain is a polymethylene group containing at least one doublebond in which one or more hydrogen atoms are replaced with asubstituent. Suitable substituents include those described below for asubstituted aliphatic group.

As used herein, the term “cyclopropylenyl” refers to a bivalentcyclopropyl group of the following structure:

As used herein, the term “cyclobutylenyl” refers to a bivalentcyclobutyl group of the following structure:

As used herein, the term “oxetanyl” refers to a bivalent oxetanyl groupof the following structure:

The term “halogen” means F, Cl, Br, or I.

The term “aryl” used alone or as part of a larger moiety as in“aralkyl,” “aralkoxy,” or “aryloxyalkyl,” refers to monocyclic orbicyclic ring systems having a total of five to fourteen ring members,wherein at least one ring in the system is aromatic and wherein eachring in the system contains 3 to 7 ring members. The term “aryl” may beused interchangeably with the term “aryl ring.”

The term “aryl” used alone or as part of a larger moiety as in“aralkyl,” “aralkoxy,” or “aryloxyalkyl,” refers to monocyclic andbicyclic ring systems having a total of five to 10 ring members, whereinat least one ring in the system is aromatic and wherein each ring in thesystem contains three to seven ring members. The term “aryl” may be usedinterchangeably with the term “aryl ring”. In certain embodiments of thepresent invention, “aryl” refers to an aromatic ring system whichincludes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl andthe like, which may bear one or more substituents. Also included withinthe scope of the term “aryl,” as it is used herein, is a group in whichan aromatic ring is fused to one or more nonaromatic rings, such asindanyl, phthalimidyl, naphthimidyl, phenanthridinyl, ortetrahydronaphthyl, and the like.

The terms “heteroaryl” and “heteroar-,” used alone or as part of alarger moiety, e.g., “heteroaralkyl,” or “heteroaralkoxy,” refer togroups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms;having 6, 10, or 14 π electrons shared in a cyclic array; and having, inaddition to carbon atoms, from one to five heteroatoms. The term“heteroatom” refers to nitrogen, oxygen, or sulfur, and includes anyoxidized form of nitrogen or sulfur, and any quaternized form of a basicnitrogen. Heteroaryl groups include, without limitation, thienyl,furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl,oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl,thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl,purinyl, naphthyridinyl, and pteridinyl. The terms “heteroaryl” and“heteroar-”, as used herein, also include groups in which aheteroaromatic ring is fused to one or more aryl, cycloaliphatic, orheterocyclyl rings, where the radical or point of attachment is on theheteroaromatic ring. Nonlimiting examples include indolyl, isoindolyl,benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl,benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl,quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl,phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Aheteroaryl group may be mono or bicyclic. The term “heteroaryl” may beused interchangeably with the terms “heteroaryl ring,” “heteroarylgroup,” or “heteroaromatic,” any of which terms include rings that areoptionally substituted. The term “heteroaralkyl” refers to an alkylgroup substituted by a heteroaryl, wherein the alkyl and heteroarylportions independently are optionally substituted.

As used herein, the terms “heterocycle,” “heterocyclyl,” “heterocyclicradical,” and “heterocyclic ring” are used interchangeably and refer toa stable 5- to 7-membered monocyclic or 7-10-membered bicyclicheterocyclic moiety that is either saturated or partially unsaturated,and having, in addition to carbon atoms, one or more, preferably one tofour, heteroatoms, as defined above. When used in reference to a ringatom of a heterocycle, the term “nitrogen” includes a substitutednitrogen. As an example, in a saturated or partially unsaturated ringhaving 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, thenitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as inpyrrolidinyl), or ⁺NR (as in N-substituted pyrrolidinyl).

A heterocyclic ring can be attached to its pendant group at anyheteroatom or carbon atom that results in a stable structure and any ofthe ring atoms can be optionally substituted. Examples of such saturatedor partially unsaturated heterocyclic radicals include, withoutlimitation, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl,piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. Theterms “heterocycle,” “heterocyclyl,” “heterocyclyl ring,” “heterocyclicgroup,” “heterocyclic moiety,” and “heterocyclic radical,” are usedinterchangeably herein, and also include groups in which a heterocyclylring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings,such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, ortetrahydroquinolinyl, where the radical or point of attachment is on theheterocyclyl ring. A heterocyclyl group may be mono- or bicyclic. Theterm “heterocyclylalkyl” refers to an alkyl group substituted by aheterocyclyl, wherein the alkyl and heterocyclyl portions independentlyare optionally substituted.

As used herein, the term “partially unsaturated” refers to a ring moietythat includes at least one double or triple bond. The term “partiallyunsaturated” is intended to encompass rings having multiple sites ofunsaturation, but is not intended to include aryl or heteroarylmoieties, as herein defined.

As described herein, compounds of the invention may contain “optionallysubstituted” moieties. In general, the term “substituted,” whetherpreceded by the term “optionally” or not, means that one or morehydrogens of the designated moiety are replaced with a suitablesubstituent. Unless otherwise indicated, an “optionally substituted”group may have a suitable substituent at each substitutable position ofthe group, and when more than one position in any given structure may besubstituted with more than one substituent selected from a specifiedgroup, the substituent may be either the same or different at everyposition. Combinations of substituents envisioned by this invention arepreferably those that result in the formation of stable or chemicallyfeasible compounds. The term “stable,” as used herein, refers tocompounds that are not substantially altered when subjected toconditions to allow for their production, detection, and, in certainembodiments, their recovery, purification, and use for one or more ofthe purposes disclosed herein.

Suitable monovalent substituents on a substitutable carbon atom of an“optionally substituted” group are independently halogen;—(CH₂)₀₋₄R^(∘); —(CH₂)₀₋₄OR^(∘); —O(CH₂)₀₋₄R^(∘), —O—(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄CH(OR^(∘))₂; —(CH₂)₀₋₄SR^(∘); —(CH₂)₀₋₄Ph, which may besubstituted with R^(∘); —(CH₂)₀₋₄O(CH₂)₀₋₁Ph which may be substitutedwith R^(∘); —CH═CHPh, which may be substituted with R^(∘);—(CH₂)₀₋₄O(CH₂)₀₋₁-pyridyl which may be substituted with R^(∘); —NO₂;—CN; —N₃; —(CH₂)₀₋₄N(R^(∘))₂; —(CH₂)₀₋₄N(R^(∘))C(O)R^(∘);—N(R^(∘))C(S)R^(∘); —(CH₂)₀₋₄N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))C(S)NR^(∘)₂; —(CH₂)₀₋₄N(R^(∘))C(O)OR^(∘); —N(R^(∘))N(R^(∘))C(O)R^(∘);—N(R^(∘))N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))N(R^(∘))C(O)OR^(∘);—(CH₂)₀₋₄C(O)R^(∘); —C(S)R^(∘); —(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄C(O)SR^(∘); —(CH₂)₀₋₄C(O)OSiR^(∘) ₃; —(CH₂)₀₋₄OC(O)R^(∘);—OC(O)(CH₂)₀₋₄SR—, SC(S)SR^(∘); —(CH₂)₀₋₄SC(O)R^(∘); —(CH₂)₀₋₄C(O)NR⁰²;—C(S)NR^(∘) ₂; —C(S)SR^(∘); —SC(S)SR^(∘), —(CH₂)₀₋₄OC(O)NR⁰²;—C(O)N(OR^(∘))R^(∘); —C(O)C(O)R^(∘); —C(O)CH₂C(O)R^(∘);—C(NOR^(∘))R^(∘); —(CH₂)₀₋₄SSR^(∘); —(CH₂)₀₋₄S(P)₂R^(∘);—(CH₂)₀₋₄S(O)₂OR^(∘); —(CH₂)₀₋₄OS(O)₂R^(∘); —S(O)₂NR^(∘) ₂;—(CH₂)₀₋₄S(O)R^(∘); —N(R^(∘))S(O)₂NR^(∘) ₂; —N(R^(∘))S(P)₂R^(∘);—N(OR^(∘))R^(∘); —C(NH)NR^(∘) ₂; —P(O)₂R^(∘); —P(O)R^(∘) ₂; —OP(O)R^(∘)₂; —OP(O)(OR^(∘))₂; SiR^(∘) ₃; —(C₁₋₄ straight or branchedalkylene)O—N(R^(∘))₂; or —(C₁₋₄ straight or branchedalkylene)C(O)O—N(R^(∘))₂, wherein each R^(∘) may be substituted asdefined below and is independently hydrogen, C₁₋₆ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, —CH₂-(5-6 membered heteroaryl ring), or a 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(∘), taken together with their intervening atom(s), form a3-12-membered saturated, partially unsaturated, or aryl mono- orbicyclic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, which may be substituted as defined below.

Suitable monovalent substituents on R^(∘) (or the ring formed by takingtwo independent occurrences of R^(∘) together with their interveningatoms), are independently halogen, —(CH₂)₀₋₂R^(●), -(haloR^(●)),—(CH₂)₀₋₂OH, —(CH₂)₀₋₂OR^(●), —(CH₂)₀₋₂CH(OR^(●))₂; —O(haloR^(●)), —CN,—N₃, —(CH₂)₀₋₂C(O)R^(●), —(CH₂)₀₋₂C(O)OH, —(CH₂)₀₋₂C(O)OR^(●),—(CH₂)₀₋₂SR^(●), —(CH₂)₀₋₂SH, —(CH₂)₀₋₂NH₂, —(CH₂)₀₋₂NHR^(●),—(CH₂)₀₋₂NR^(●) ₂, —NO₂, —SiR^(●) ₃, —OSiR^(●) ₃, —C(O)SR^(●), —(C₁₋₄straight or branched alkylene)C(O)OR^(●), or —SSR^(●) wherein each R^(●)is unsubstituted or where preceded by “halo” is substituted only withone or more halogens, and is independently selected from C₁₋₄ aliphatic,—CH₂Ph, —O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. Suitable divalent substituents on asaturated carbon atom of R^(∘) include ═O and ═S.

Suitable divalent substituents on a saturated carbon atom of an“optionally substituted” group include the following: ═O, ═S, ═NNR*₂,═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O)₂R*, ═NR*, ═NOR*, —O(C(R*₂))₂₋₃O—, or—S(C(R*₂))₂₋₃S—, wherein each independent occurrence of R^(●) isselected from hydrogen, C₁₋₆ aliphatic which may be substituted asdefined below, or an unsubstituted 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. Suitable divalent substituents thatare bound to vicinal substitutable carbons of an “optionallysubstituted” group include: —O(CR*₂)₂₋₃O—, wherein each independentoccurrence of R* is selected from hydrogen, C₁₋₆ aliphatic which may besubstituted as defined below, or an unsubstituted 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R* include halogen,—R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN, —C(O)OH,—C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or —NO₂, wherein each R^(●) isunsubstituted or where preceded by “halo” is substituted only with oneor more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

Suitable substituents on a substitutable nitrogen of an “optionallysubstituted” group include —R^(†), —NR^(†) ₂, —C(O)R^(†), —C(O)OR^(†),—C(O)C(O)R^(†), —C(O)CH₂C(O)R^(†), —S(O)₂R^(†), —S(O)₂NR^(†) ₂,—C(S)NR^(†) ₂, —C(NH)NR^(†) ₂, or —N(R^(†))S(O)₂R^(†); wherein each R isindependently hydrogen, C₁₋₆ aliphatic which may be substituted asdefined below, unsubstituted —OPh, or an unsubstituted 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(†), taken together with their intervening atom(s) form anunsubstituted 3-12-membered saturated, partially unsaturated, or arylmono- or bicyclic ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R^(†) are independentlyhalogen, —R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN,—C(O)OH, —C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or —NO₂, wherein eachR^(●) is unsubstituted or where preceded by “halo” is substituted onlywith one or more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge etal., describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein byreference. Pharmaceutically acceptable salts of the compounds of thisinvention include those derived from suitable inorganic and organicacids and bases. Examples of pharmaceutically acceptable, nontoxic acidaddition salts are salts of an amino group formed with inorganic acidssuch as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuricacid and perchloric acid or with organic acids such as acetic acid,oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid ormalonic acid or by using other methods used in the art such as ionexchange. Other pharmaceutically acceptable salts include adipate,alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate,borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate,propionate, stearate, succinate, sulfate, tartrate, thiocyanate,p-toluenesulfonate, undecanoate, valerate salts, and the like.

Salts derived from appropriate bases include alkali metal, alkalineearth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. Representative alkali oralkaline earth metal salts include sodium, lithium, potassium, calcium,magnesium, and the like. Further pharmaceutically acceptable saltsinclude, when appropriate, nontoxic ammonium, quaternary ammonium, andamine cations formed using counterions such as halide, hydroxide,carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and arylsulfonate.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, Z and E double bond isomers,and Z and E conformational isomers. Therefore, single stereochemicalisomers as well as enantiomeric, diastereomeric, and geometric (orconformational) mixtures of the present compounds are within the scopeof the invention. Unless otherwise stated, all tautomeric forms of thecompounds of the invention are within the scope of the invention.Additionally, unless otherwise stated, structures depicted herein arealso meant to include compounds that differ only in the presence of oneor more isotopically enriched atoms. For example, compounds having thepresent structures including the replacement of hydrogen by deuterium ortritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enriched carbonare within the scope of this invention. Such compounds are useful, forexample, as analytical tools, as probes in biological assays, or astherapeutic agents in accordance with the present invention.

3. Description of Exemplary Embodiments

In certain embodiments, the present invention provides inhibitors ofAHR. In some embodiments, such compounds include those of formula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   Ring A is selected from:

-   each R¹ is independently selected from R, —C(O)R, —C(O)OR, —SO₂R,    —C(O)N(R)₂, or —SO₂RN(R)₂;-   each R is independently hydrogen, deuterium, or an optionally    substituted group selected from C₁₋₆ aliphatic, a 3-8 membered    saturated or partially unsaturated monocyclic carbocyclic ring,    phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring; a 4-8    membered saturated or partially unsaturated monocyclic heterocyclic    ring having 1-2 heteroatoms independently selected from nitrogen,    oxygen, or sulfur, a 5-6 membered monocyclic heteroaromatic ring    having 1-4 heteroatoms independently selected from nitrogen, oxygen,    or sulfur, or an 8-10 membered bicyclic heteroaromatic ring having    1-5 heteroatoms independently selected from nitrogen, oxygen, or    sulfur;-   each or R^(x), R^(y), and R^(z) is independently selected from R,    halogen, cyano, nitro, —OR, —SR, —N(R)₂, —N(R)C(O)R, —C(O)N(R)₂,    —N(R)C(O)N(R)₂, —N(R)C(O)OR, —OC(O)N(R)₂, —N(R)SO₂R, —SO₂RN(R)₂,    —C(O)R, —C(O)OR, —OC(O)R, —C(O)OR, —S(O)R, or —SO₂R, or:

two R^(x) on the same carbon are taken together to form ═O or ═S; or:

two R^(y) on the same carbon are taken together to form ═O or ═S;

-   each of m and n is independently 1, 2, 3, 4, or 5;-   Ring B is phenyl, a 5-6 membered monocyclic heteroaromatic ring    having 1-4 heteroatoms independently selected from nitrogen, oxygen,    or sulfur, or an 8-10 membered bicyclic heteroaryl ring having 1-4    heteroatoms independently selected from nitrogen, oxygen, or sulfur;-   Ring C is phenyl or a 5-6 membered heteroaryl ring having 1-4    heteroatoms independently selected from nitrogen, oxygen, or sulfur;    or an 8-10 membered bicyclic heteroaryl ring having 1-4 heteroatoms    independently selected from nitrogen, oxygen, or sulfur;-   L¹ is a covalent bond or an optionally substituted C₁₋₆ membered    straight or branched bivalent hydrocarbon chain wherein a methylene    unit of L¹ is optionally replaced with -Cy-, —O—, —S—, —NR—, —C(O)—,    —C(O)O—, —OC(O)—, —C(O)N(R)—, —N(R)C(O)—, —SO₂—, —N(R)SO₂—, or    SO₂N(R)—S; and-   -Cy- is a 3-8 membered bivalent saturated, partially unsaturated, or    aromatic monocyclic ring having 0-2 heteroatoms independently    selected from nitrogen, oxygen, or sulfur, or an 8-10 membered    bivalent saturated, partially unsaturated, or aromatic bicyclic ring    having 0-3 heteroatoms independently selected from nitrogen, oxygen,    or sulfur.

In some embodiments, the present invention provides inhibitors of AHR,such compounds include include those of formula I′:

or a pharmaceutically acceptable salt thereof, wherein:

-   Ring A is selected from:

-   each p is independently 0, 1, or 2, as valency will allow;-   each R¹ is independently selected from R, —C(O)R, —C(O)OR, —SO₂R,    —C(O)N(R)₂, or —SO₂RN(R)₂;-   each R is independently hydrogen, deuterium, or an optionally    substituted group selected from C₁₋₆ aliphatic, a 3-8 membered    saturated or partially unsaturated monocyclic carbocyclic ring,    phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring; a 4-8    membered saturated or partially unsaturated monocyclic heterocyclic    ring having 1-2 heteroatoms independently selected from nitrogen,    oxygen, or sulfur, a 5-6 membered monocyclic heteroaromatic ring    having 1-4 heteroatoms independently selected from nitrogen, oxygen,    or sulfur, or an 8-10 membered bicyclic heteroaromatic ring having    1-5 heteroatoms independently selected from nitrogen, oxygen, or    sulfur; or two R on the same nitrogen are taken together with their    intervening atoms to form a 4-7 membered saturated, partially    unsaturated, or aromatic ring having 1-2 heteroatoms in addition to    the nitrogen independently selected from oxygen, nitrogen, or    sulfur;-   each or R^(x), R^(y), and R^(z) is independently selected from R,    halogen, cyano, nitro, —OR, —SR, —N(R)₂, —N(R)C(O)R, —C(O)N(R)₂,    —C(O)N(R)OR, —N(R)C(O)N(R)₂, —N(R)C(O)OR, —OC(O)N(R)₂, —N(R)SO₂R,    —SO₂RN(R)₂, —C(O)R, —C(O)OR, —OC(O)R, —C(O)OR, —S(O)R, or —SO₂R, or:

two R^(x) on the same carbon are taken together to form ═O or ═S; or:

two R^(y) on the same carbon are taken together to form ═O or ═S;

-   each of m and n is independently 1, 2, 3, 4, or 5;-   Ring B is phenyl, a 7-10 membered bicyclic partially unsaturated or    aromatic carbocyclic ring, a 5-6 membered monocyclic heteroaromatic    ring having 1-4 heteroatoms independently selected from nitrogen,    oxygen, or sulfur, an 8-10 membered bicyclic heteroaryl ring having    1-4 heteroatoms independently selected from nitrogen, oxygen, or    sulfur, or a 12-15 membered partially unsaturated or aromatic    tricyclic ring having 1-4 heteroatoms independently selected from    nitrogen, oxygen, or sulfur;-   Ring C is phenyl or a 5-6 membered heteroaryl ring having 1-4    heteroatoms independently selected from nitrogen, oxygen, or sulfur;-   L¹ is a covalent bond or an optionally substituted C₁₋₆ membered    straight or branched bivalent hydrocarbon chain wherein a methylene    unit of L¹ is optionally replaced with -Cy-, —O—, —S—, —NR—, —C(O)—,    —C(O)O—, —OC(O)—, —C(O)N(R)—, —N(R)C(O)—, —SO₂—, —N(R)SO₂—, or    SO₂N(R)—S; and-   Cy- is a 3-8 membered bivalent saturated, partially unsaturated, or    aromatic monocyclic ring having 0-2 heteroatoms independently    selected from nitrogen, oxygen, or sulfur, or an 8-10 membered    bivalent saturated, partially unsaturated, or aromatic bicyclic ring    having 0-3 heteroatoms independently selected from nitrogen, oxygen,    or sulfur.

In some embodiments, the present invention provides a compound offormula I or formula I′, with the proviso that when Ring A is

Ring B is not

and/or Ring C is not

and/or R¹ is not

In some embodiments, the present invention provides a compound offormula I or formula I′, with the proviso that L¹ is not —NHCH₂CH₂—. Insome embodiments, the present invention provides a compound of formula Ior formula I′, with the proviso that when Ring A is

L¹ is not —NHCH₂CH₂—.

In some embodiments, the present invention provides a compound offormula I or formula I′, with the proviso that the compound is otherthan:

In some embodiments, a provided compound is other than

In some embodiments, a provided compound is other than

In some embodiments, a provided compound is other than

In some embodiments, a provided compound is other than

In some embodiments, a provided compound is other than

In some embodiments, a provided compound is other than

As defined generally above, R¹ is R, —C(O)R, —C(O)OR, —SO₂R, —C(O)N(R)₂,or —SO₂RN(R)₂. In some embodiments, R¹ is hydrogen. In some embodiments,R¹ is R. In some embodiments, R¹ is —C(O)R. In some embodiments, R¹ is—C(O)OR. In some embodiments, R¹ is —SO₂R. In some embodiments, R¹ is—C(O)N(R)₂. In some embodiments, R¹ is —SO₂RN(R)₂. In some embodiments,R¹ is hydrogen. In some embodiments, R¹ is deuterium. In someembodiments, R¹ is an optionally substituted group selected from C₁₋₆aliphatic. In some embodiments, R¹ is selected from those depicted inTable 1, below.

As defined generally above, each R^(x) is independently R, halogen,cyano, nitro, —OR, —SR, —N(R)₂, —N(R)C(O)R, —C(O)N(R)₂, —C(O)N(R) OR,—N(R)C(O)N(R)₂, —N(R)C(O)OR, —OC(O)N(R)₂, —N(R)SO₂R, —SO₂RN(R)₂, —C(O)R,—C(O)OR, —OC(O)R, —C(O)OR, —S(O)R, or —SO₂R, or two R^(x) on the samecarbon are taken together to form ═O or ═S. In some embodiments, eachR^(x) is the same. In some embodiments, each R^(x) is different. In someembodiments, R^(x) is hydrogen. In some embodiments, R^(x) is R. In someembodiments, R^(x) is halogen. In some embodiments, R^(x) is cyano. Insome embodiments, R^(x) is nitro. In some embodiments, IV is —OR. Insome embodiments, R^(x) is —SR. In some embodiments, R^(x) is —N(R)₂. Insome embodiments, R^(x) is —N(R)C(O)R. In some embodiments, R^(x) is—C(O)N(R)₂. In some embodiments, R^(x) is —C(O)N(R)OR. In someembodiments, R^(x) is —N(R)C(O)N(R)₂. In some embodiments, R^(x) is—N(R)C(O)OR. In some embodiments, R^(x) is —OC(O)N(R)₂. In someembodiments, R^(x) is —N(R)SO₂R. In some embodiments, R^(x) is—SO₂RN(R)₂. In some embodiments, R^(x) is —C(O)R. In some embodiments,R^(x) is —C(O)OR. In some embodiments, R^(x) is CO(O)R. In someembodiments, R^(x) is —S(O)R. In some embodiments, R^(x) is —SO₂R. Insome embodiments, two IV on the same carbon are taken together to form═O or ═S. In some embodiments, R^(x) is hydrogen. In some embodiments,R^(x) is deuterium. In some embodiments, R^(x) is an optionallysubstituted group selected from C₁₋₆ aliphatic. In some embodiments,R^(x) is selected from those depicted in Table 1, below.

As defined generally above, each R^(y) is independently R, halogen,cyano, nitro, —OR, —SR, —N(R)₂, —N(R)C(O)R, —C(O)N(R)₂, —C(O)N(R)OR,—N(R)C(O)N(R)₂, —N(R)C(O)OR, —OC(O)N(R)₂, —N(R)SO₂R, —SO₂RN(R)₂, —C(O)R,—C(O)OR, —OC(O)R, —C(O)OR, —S(O)R, or —SO₂R, or two R^(y) on the samecarbon are taken together to form ═O or ═S. In some embodiments, eachR^(y) is the same. In some embodiments, each R^(y) is different. In someembodiments, R^(y) is hydrogen. In some embodiments, R^(y) is R. In someembodiments, R^(y) is halogen. In some embodiments, R^(y) is cyano. Insome embodiments, R^(y) is nitro. In some embodiments, R^(y) is —OR. Insome embodiments, R^(y) is —SR. In some embodiments, R^(y) is —N(R)₂. Insome embodiments, R^(y) is —C(O)N(R)OR. In some embodiments, R^(y) is—N(R)C(O)R. In some embodiments, R^(y) is —C(O)N(R)₂. In someembodiments, R^(y) is —N(R)C(O)N(R)₂. In some embodiments, R^(y) is—N(R)C(O)OR. In some embodiments, R^(y) is —OC(O)N(R)₂. In someembodiments, R^(y) is —N(R)SO₂R. In some embodiments, R^(y) is—SO₂RN(R)₂. In some embodiments, R^(y) is —C(O)R. In some embodiments,R^(y) is —C(O)OR. In some embodiments, R^(y) is CO(O)R. In someembodiments, R^(y) is —S(O)R. In some embodiments, R^(y) is —SO₂R. Insome embodiments, two R^(y) on the same carbon are taken together toform ═O or ═S. In some embodiments, R^(y) is hydrogen. In someembodiments, R^(y) is deuterium. In some embodiments, R^(y) is anoptionally substituted group selected from C₁₋₆ aliphatic. In someembodiments, R^(y) is selected from those depicted in Table 1, below.

As defined generally above, each R^(z) is independently R, halogen,cyano, nitro, —OR, —SR, —N(R)₂, —N(R)C(O)R, —C(O)N(R)₂, —C(O)N(R)OR,—N(R)C(O)N(R)₂, —N(R)C(O)OR, —OC(O)N(R)₂, —N(R)SO₂R, —SO₂RN(R)₂, —C(O)R,—C(O)OR, —OC(O)R, —C(O)OR, —S(O)R, or —SO₂R. In some embodiments, R^(z)is hydrogen. In some embodiments, R^(z) is R. In some embodiments, R^(z)is halogen. In some embodiments, R^(z) is cyano. In some embodiments,R^(z) is nitro. In some embodiments, R^(z) is —OR. In some embodiments,R^(z) is —SR. In some embodiments, R^(z) is —N(R)₂. In some embodiments,R^(z) is —C(O)N(R)OR. In some embodiments, R^(z) is N(R)C(O)R. In someembodiments, R^(z) is —C(O)N(R)₂. In some embodiments, R^(z) isN(R)C(O)N(R)₂. In some embodiments, R^(z) is —N(R)C(O)OR. In someembodiments, R^(z) is OC(O)N(R)₂. In some embodiments, R^(z) is—N(R)SO₂R. In some embodiments, R^(z) is —SO₂RN(R)₂. In someembodiments, R^(z) is —C(O)R. In some embodiments, R^(z) is —C(O)OR. Insome embodiments, R^(z) is CO(O)R. In some embodiments, R^(z) is —S(O)R.In some embodiments, R^(z) is —SO₂R. In some embodiments, R^(z) ishydrogen. In some embodiments, R^(z) is deuterium. In some embodiments,R^(z) is an optionally substituted group selected from C₁₋₆ aliphatic.In some embodiments, R^(z) is selected from those depicted in Table 1,below.

As defined generally above, p is 0, 1 or 2. In some embodiments, p is 0.In some embodiments, p is 1. In some embodiments, p is 2. In someembodiments, p is selected from those depicted in Table 1, below.

As defined generally above, n is 1, 2, 3, 4 or 5. In some embodiments, nis 1. In some embodiments, n is 2. In some embodiments, n is 3. In someembodiments, n is 4. In some embodiments, n is 5. In some embodiments, nis selected from those depicted in Table 1, below.

As defined generally above, m is 1, 2, 3, 4 or 5. In some embodiments, mis 1. In some embodiments, m is 2. In some embodiments, m is 3. In someembodiments, m is 4. In some embodiments, m is 5. In some embodiments, mis selected from those depicted in Table 1, below.

As defined generally above, Ring B is phenyl, a 7-10 membered bicyclicpartially unsaturated or aromatic carbocyclic ring, a 5-6 memberedmonocyclic heteroaromatic ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, an 8-10 membered bicyclicheteroaryl ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, or a 12-15 membered partially unsaturatedor aromatic tricyclic ring having 1-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. In some embodiments, Ring B is a 7-10membered bicyclic partially unsaturated or aromatic carbocyclic ring. Insome embodiments, Ring B is a 12-15 membered partially unsaturated oraromatic tricyclic ring having 1-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. In some embodiments, Ring B is a 5-6membered monocyclic heteroaromatic ring having 1-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur. In someembodiments, Ring B is an 8-10 membered bicyclic heteroaryl ring having1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.In some embodiments, Ring B is phenyl. In some embodiments, Ring B isthienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl,tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl,thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl,purinyl, naphthyridinyl or pteridinyl, indolycarl, isoindolyl,benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl,benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl,quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl,phenazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, orpyrido[2,3-b]-1,4-oxazin-3(4H)-one. In some embodiments, Ring B isselected from those depicted in Table 1, below.

As defined generally above, Ring C is phenyl, a 5-6 membered monocyclicheteroaromatic ring having 1-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, or an 8-10 membered bicyclic heteroarylring having 1-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur. In some embodiments, Ring C is phenyl. In someembodiments, Ring C is a 5-6 membered heteroaryl ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur. Insome embodiments, Ring C is thienyl, furanyl, pyrrolyl, imidazolyl,pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl,thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl,pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl orpteridinyl, indolyl, isoindolyl, benzothienyl, benzofuranyl,dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl,isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl,4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl, orpyrido[2,3-b]-1,4-oxazin-3(4H)-one. In some embodiments, Ring C isselected from those depicted in Table 1, below.

As defined generally above, L′ is a covalent bond or an optionallysubstituted C₁₋₆ membered straight or branched bivalent hydrocarbonchain wherein a methylene unit of L¹ is optionally replaced with -Cy-,—O—, —S—, —NR—, —C(O)—, —C(O)O—, —OC(O)—, —C(O)N(R)—, —N(R)C(O)—, —SO₂—,—N(R)SO₂—, or SO₂N(R)—S. In some embodiments, L¹ is a covalent bond. Insome embodiments, L¹ is an optionally substituted C₁₋₆ membered straightor branched bivalent hydrocarbon chain. In some embodiments, L¹ is -Cy-.In some embodiments, L¹ is phenylene, heterocyclylene, heteroarylene,cyclopropylene, cyclobutylenyl, cyclopentylene, cyclohexylene oroxetanyl. In some embodiments, L¹ is —NR—. In some embodiments, L¹ isN(CH₂)₂—. In some embodiments, L¹ is selected from those depicted inTable 1, below.

In some embodiments, -Cy- is phenylene, heterocyclylene, heteroarylene,cyclopropylene, cyclobutylenyl, cyclopentylene, cyclohexylene andoxetanyl. In some embodiments, -Cy- is selected from:

wherein X is a heteroatom selected from nitrogen, oxygen, or sulfur. Insome embodiments, -Cy- is selected from those depicted in Table 1,below.

In some embodiments, the present invention provides a compound selectedfrom any of formulae I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-i, I-j,I-k, I-l, I-m, I-n, I-o and I-p:

or a pharmaceutically acceptable salt thereof; wherein each variable isas defined herein and described in embodiments for formula I and formulaI′, supra, or described in embodiments herein, both singly and incombination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae II-a, II-b, II-c, II-d, II-e, II-f, II-g, II-h,II-j, II-k, II-l, II-m, II-n, II-o and II-p

or a pharmaceutically acceptable salt thereof; wherein each variable isas defined herein and described in embodiments for formula I and formulaI′, supra, or described in embodiments herein, both singly and incombination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae III-a, III-b, III-c, III-d, III-e, III-f, III-g,III-h, III-i, III-j, III-k, III-l, III-m, III-n, III-o, III-p, III-q,III-r, III-s, III-t, III-u, III-v, III-w, III-x, III-y, III-z, III-aa,III-bb, III-cc, III-dd, III-ee, III-ff, III-gg, III-hh, III-ii, III-jjand III-kk:

or a pharmaceutically acceptable salt thereof; wherein each variable isas defined herein and described in embodiments for formula I and formulaI′, supra, or described in embodiments herein, both singly and incombination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae IV-a, IV-b, IV-c, IV-d, IV-e, IV-f, IV-g, IV-h,IV-i, IV-j, IV-k, IV-l, IV-m, IV-n, IV-o, IV-p and IV-q:

or a pharmaceutically acceptable salt thereof; wherein each variable isas defined herein and described in embodiments for formula I and formulaI′, supra, or described in embodiments herein, both singly and incombination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae V-a, V-b, V-c, V-d, V-e, V-f, V-g, V-h, V-i, V-j,V-k and V-l:

or a pharmaceutically acceptable salt thereof; wherein each variable isas defined herein and described in embodiments for formula I and formulaI′, supra, or described in embodiments herein, both singly and incombination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae VI-a, VI-b, VI-c, VI-d, VI-e and VI-f:

or a pharmaceutically acceptable salt thereof wherein X is N or CH;wherein each variable is as defined herein and described in embodimentsfor formula I and formula I′, supra, or described in embodiments herein,both singly and in combination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae VII-a, VII-b, VII-c, VII-d, VII-e, VII-f and VII-g:

or a pharmaceutically acceptable salt thereof wherein X is N or CH;wherein each variable is as defined herein and described in embodimentsfor formula I and formula I′, supra, or described in embodiments herein,both singly and in combination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae VIII-a, VIII-b, VIII-c, VIII-d, VIII-e and VIII-f:

or a pharmaceutically acceptable salt thereof wherein X is N or CH;wherein each variable is as defined herein and described in embodimentsfor formula I and formula I′, supra, or described in embodiments herein,both singly and in combination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae IX-a, IX-b and IX-c:

or a pharmaceutically acceptable salt thereof wherein X is N or CH;wherein each variable is as defined herein and described in embodimentsfor formula I and formula I′, supra, or described in embodiments herein,both singly and in combination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae X-a, X-b, X-c, X-d, X-e, X-f, X-g, X-h and X-i:

or a pharmaceutically acceptable salt thereof wherein X is N or CH;wherein each variable is as defined herein and described in embodimentsfor formula I and formula I′, supra, or described in embodiments herein,both singly and in combination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae XI-a, XI-b, XI-c, XI-d, XI-e, XI-f, XI-g, XI-h andXI-i:

or a pharmaceutically acceptable salt thereof wherein X is N or CH;wherein each variable is as defined herein and described in embodimentsfor formula I and formula I′, supra, or described in embodiments herein,both singly and in combination.

In some embodiments, the present invention provides a compound selectedfrom any of formulae XII-a, XII-b and XI-c:

or a pharmaceutically acceptable salt thereof wherein X is N or CH;wherein each variable is as defined herein and described in embodimentsfor formula I and formula I′, supra, or described in embodiments herein,both singly and in combination.

In some embodiments, the present invention provides a compound offormula I and formula I′ provided that when Ring A is

Ring B is not

and/or Ring C is not

and/or R¹ is not

In some embodiments, the present invention provides a compound offormula I and formula I′ provided that L¹ is not —NHCH₂CH₂—. In someembodiments, the present invention provides a compound of formula I andformula I′ provided that when Ring A is

L¹ is not —NHCH₂CH₂—.

Exemplary compounds of the present invention are set forth in Table 1,below:

TABLE 1 Exemplary Compounds of Formula I I-1

I-2

I-3

I-4

I-5

I-6

I-7

I-8

I-9

I-10

I-11

I-12

I-13

I-14

I-15

I-16

I-17

I-18

I-19

I-20

I-21

I-22

I-23

I-24

I-25

I-26

I-27a

I-27b

I-28a

I-28b

I-29

I-30

I-31

I-32a

I-32b

I-33

I-34

I-35

I-36

I-37a

I-37b

I-38

I-39

I-40

I-41

I-42a

I-42b

I-43

I-44

I-45

I-46

I-47

I-48

I-49

I-50

I-51

I-52

I-53

I-54

I-55

I-56

I-57

I-58

I-59

I-60

I-61

I-62

I-63

I-64

I-65

I-66

I-67

I-68

I-69

I-70

I-71

I-72

I-73

I-74

I-75a

I-75b

I-75c

I-76

I-77

I-78

I-79

I-80

I-81

I-82a

I-82b

I-82c

I-83

I-84

I-85

I-86

I-87

I-88

I-89

I-90

I-91

I-92

I-93

I-94a

I-94b

I-95

I-96

I-97a

I-97b

I-98a

I-98b

I-99

I-100

I-101

I-102

I-103

I-104

I-105

I-106

I-107

I-108

I-109

I-110

I-111

I-112

I-113

I-114

I-115

I-116

I-117

I-118

I-119

I-120

I-121

I-122

I-123

I-124

I-125

I-126

I-128a

I-128b

I-129

I-130

I-131

I-132

I-133

I-134

I-135

I-136

I-137a

I-137b

I-137c

I-139a

I-139b

I-139c

I-141

I-142

I-143

I-144a

I-144b

I-146a

I-146b

I-148

I-149

I-150a

I-150b

I-152a

I-152b

I-152c

I-154

I-155

I-156

I-157

I-158a

I-158b

I-160a

I-160b

I-160c

I-162

I-163

I-164

I-165

I-166

I-167

I-168

I-169

I-170

I-171

I-172

I-173

I-174

I-175

I-176

I-177

I-178

I-179

I-180

I-181

I-182

I-183

I-184

I-185

I-186

I-187a

I-187b

I-189

I-190a

I-190b

I-192

I-193

I-194

I-195

I-196

I-197

I-198a

I-198b

I-198c

I-200

I-201

I-202

I-203

I-204a

I-204b

I-206

I-207

I-208

I-209

I-210

I-211

I-212

I-213

I-214

I-215

I-216a

I-216b

I-218

I-219

I-220

I-221

I-222a

I-222b

I-222c

I-224

I-225

I-226

I-227

I-228

I-229

I-230

I-231

I-232

I-233

I-234

I-235

I-236

I-237

I-238

I-239

I-240

I-241

I-242

I-243a

I-243b

I-245

I-246

I-247a

I-247b

I-249a

I-249b

I-249c

I-250

I-251

I-252a

I-252b

I-252c

I-253

I-254

I-255

I-256

I-257

I-258

I-259a

I-259b

I-259c

I-260

I-261

I-262

I-263a

I-263b

I-263c

I-264

I-265

I-266

I-267

I-268

I-269

I-270

I-271

I-272

I-273

I-274

I-275

I-276

I-277

I-278

I-279

I-280

I-281

I-282

I-283

I-284

I-285

I-286

I-287

I-288

I-289

In certain embodiments, the present invention provides any compoundselected from those depicted in Table 1, above, or a pharmaceuticallyacceptable salt thereof.

4. Uses, Formulation and Administration and Pharmaceutically AcceptableCompositions

According to another embodiment, the invention provides a compositioncomprising a compound of this invention or a pharmaceutically acceptablesalt, ester, or salt of ester thereof and a pharmaceutically acceptablecarrier, adjuvant, or vehicle. The amount of compound in compositions ofthis invention is such that is effective to measurably inhibit AHR, in abiological sample or in a patient. In certain embodiments, the amount ofcompound in compositions of this invention is such that is effective tomeasurably inhibit AHR, in a biological sample or in a patient. Incertain embodiments, a composition of this invention is formulated foradministration to a patient in need of such composition. In someembodiments, a composition of this invention is formulated for oraladministration to a patient.

The term “patient,” as used herein, means an animal, preferably amammal, and most preferably a human.

The term “pharmaceutically acceptable carrier, adjuvant, or vehicle”refers to a non-toxic carrier, adjuvant, or vehicle that does notdestroy the pharmacological activity of the compound with which it isformulated. Pharmaceutically acceptable carriers, adjuvants or vehiclesthat may be used in the compositions of this invention include, but arenot limited to, ion exchangers, alumina, aluminum stearate, lecithin,serum proteins, such as human serum albumin, buffer substances such asphosphates, glycine, sorbic acid, potassium sorbate, partial glyceridemixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, sodium carboxymethylcellulose,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,polyethylene glycol and wool fat.

A “pharmaceutically acceptable derivative” means any non-toxic salt,ester, salt of an ester or other derivative of a compound of thisinvention that, upon administration to a recipient, is capable ofproviding, either directly or indirectly, a compound of this inventionor an inhibitorily active metabolite or residue thereof.

Compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously. Sterile injectable forms of thecompositions of this invention may be aqueous or oleaginous suspension.These suspensions may be formulated according to techniques known in theart using suitable dispersing or wetting agents and suspending agents.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally acceptable diluent orsolvent, for example as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or di-glycerides. Fatty acids, such as oleic acid andits glyceride derivatives are useful in the preparation of injectables,as are natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, such as carboxymethyl cellulose or similar dispersingagents that are commonly used in the formulation of pharmaceuticallyacceptable dosage forms including emulsions and suspensions. Othercommonly used surfactants, such as Tweens, Spans and other emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms may also be used for the purposes of formulation.

Pharmaceutically acceptable compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include lactose andcorn starch. Lubricating agents, such as magnesium stearate, are alsotypically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, pharmaceutically acceptable compositions of thisinvention may be administered in the form of suppositories for rectaladministration. These can be prepared by mixing the agent with asuitable non-irritating excipient that is solid at room temperature butliquid at rectal temperature and therefore will melt in the rectum torelease the drug. Such materials include cocoa butter, beeswax andpolyethylene glycols.

Pharmaceutically acceptable compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, provided pharmaceutically acceptablecompositions may be formulated in a suitable ointment containing theactive component suspended or dissolved in one or more carriers.Carriers for topical administration of compounds of this inventioninclude, but are not limited to, mineral oil, liquid petrolatum, whitepetrolatum, propylene glycol, polyoxyethylene, polyoxypropylenecompound, emulsifying wax and water. Alternatively, providedpharmaceutically acceptable compositions can be formulated in a suitablelotion or cream containing the active components suspended or dissolvedin one or more pharmaceutically acceptable carriers. Suitable carriersinclude, but are not limited to, mineral oil, sorbitan monostearate,polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol,benzyl alcohol and water.

For ophthalmic use, provided pharmaceutically acceptable compositionsmay be formulated as micronized suspensions in isotonic, pH adjustedsterile saline, or, preferably, as solutions in isotonic, pH adjustedsterile saline, either with or without a preservative such asbenzylalkonium chloride. Alternatively, for ophthalmic uses, thepharmaceutically acceptable compositions may be formulated in anointment such as petrolatum.

Pharmaceutically acceptable compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

Most preferably, pharmaceutically acceptable compositions of thisinvention are formulated for oral administration. Such formulations maybe administered with or without food. In some embodiments,pharmaceutically acceptable compositions of this invention areadministered without food. In other embodiments, pharmaceuticallyacceptable compositions of this invention are administered with food.

The amount of compounds of the present invention that may be combinedwith the carrier materials to produce a composition in a single dosageform will vary depending upon the host treated, the particular mode ofadministration. Preferably, provided compositions should be formulatedso that a dosage of between 0.01-100 mg/kg body weight/day of theinhibitor can be administered to a patient receiving these compositions.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of a compound of the present invention in the composition willalso depend upon the particular compound in the composition.

Uses of Compounds and Pharmaceutically Acceptable Compositions

The activity of a compound utilized in this invention as an inhibitor ofAHR may be assayed in vitro or in vivo. An in vivo assessment of theefficacy of the compounds of the invention may be made using an animalmodel of obesity or metabolic syndrome, e.g., a rodent or primate model.Cell-based assays may be performed using, e.g., a cell line isolatedfrom a tissue that expresses AHR. Additionally, biochemical ormechanism-based assays, e.g., transcription assays using a purifiedprotein, Northern blot, RT-PCR, etc., may be performed. In vitro assaysinclude assays that determine cell morphology, protein expression,and/or the cytotoxicity, enzyme inhibitory activity, and/or thesubsequent functional consequences of treatment of cells with compoundsof the invention. Alternate in vitro assays quantitate the ability ofthe inhibitor to bind to protein or nucleic acid molecules within thecell. Inhibitor binding may be measured by radiolabelling the inhibitorprior to binding, isolating the inhibitor/target molecule complex anddetermining the amount of radiolabel bound. Alternatively, inhibitorbinding may be determined by running a competition experiment where newinhibitors are incubated with purified proteins or nucleic acids boundto known radioligands. Detailed conditions for assaying a compoundutilized in this invention as an inhibitor of AHR are set forth in theExamples below. The aforementioned assays are exemplary and not intendedto limit the scope of the invention. The skilled practitioner canappreciate that modifications can be made to conventional assays todevelop equivalent assays that obtain the same result.

As used herein, the terms “treatment,” “treat,” and “treating” refer toreversing, alleviating, delaying the onset of, or inhibiting theprogress of a disease or disorder, or one or more symptoms thereof, asdescribed herein. In some embodiments, treatment may be administeredafter one or more symptoms have developed. In other embodiments,treatment may be administered in the absence of symptoms. For example,treatment may be administered to a susceptible individual prior to theonset of symptoms (e.g., in light of a history of symptoms and/or inlight of genetic or other susceptibility factors). Treatment may also becontinued after symptoms have resolved, for example to prevent or delaytheir recurrence.

The compounds and compositions, according to the method of the presentinvention, may be administered using any amount and any route ofadministration effective for treating or lessening the severity of ametabolic disorder or condition, cancer, a bacterial infection, a fungalinfection, a parasitic infection (e.g. malaria), an autoimmune disorder,a neurodegenerative or neurological disorder, schizophrenia, abone-related disorder, liver disease, or a cardiac disorder.

In some embodiments, the compounds and compositions, according to themethod of the present invention, may be administered using any amountand any route of administration effective for treating or lessening theseverity of a disease associated with AHR.

The exact amount required will vary from subject to subject, dependingon the species, age, and general condition of the subject, the severityof the infection, the particular agent, its mode of administration, andthe like. The compounds of the invention are preferably formulated indosage unit form for ease of administration and uniformity of dosage.The expression “dosage unit form” as used herein refers to a physicallydiscrete unit of agent appropriate for the patient to be treated. Itwill be understood, however, that the total daily usage of the compoundsand compositions of the present invention will be decided by theattending physician within the scope of sound medical judgment. Thespecific effective dose level for any particular patient or organismwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The term “patient”, as usedherein, means an animal, preferably a mammal, and most preferably ahuman.

The pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated. Incertain embodiments, the compounds of the invention may be administeredorally or parenterally at dosage levels of about 0.01 mg/kg to about 50mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subjectbody weight per day, one or more times a day, to obtain the desiredtherapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, and eye drops are also contemplatedas being within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

Uses and Methods of Treatment

According to one embodiment, the invention relates to a method ofinhibiting AHR in a biological sample comprising the step of contactingsaid biological sample with a compound of this invention, or acomposition comprising said compound.

The term “biological sample”, as used herein, includes, withoutlimitation, cell cultures or extracts thereof; biopsied materialobtained from a mammal or extracts thereof; and blood, saliva, urine,feces, semen, tears, or other body fluids or extracts thereof.

Inhibition of enzymes in a biological sample is useful for a variety ofpurposes that are known to one of skill in the art. Examples of suchpurposes include, but are not limited to biological assays, geneexpression studies, and biological target identification.

Another embodiment of the present invention relates to a method ofinhibiting AHR in a patient comprising the step of administering to saidpatient a compound of the present invention, or a composition comprisingsaid compound.

Provided compounds are inhibitors of AHR and are therefore useful fortreating one or more disorders associated with activity of AHR. Thus, incertain embodiments, the present invention provides a method fortreating an AHR-mediated disorder comprising the step of administeringto a patient in need thereof a compound of the present invention, orpharmaceutically acceptable composition thereof.

As used herein, the terms “AHR-mediated” disorders, diseases, and/orconditions as used herein means any disease or other deleteriouscondition in which AHR, or a mutant thereof, are known to play a role.Accordingly, another embodiment of the present invention relates totreating or lessening the severity of one or more diseases in which AHR,or a mutant thereof, are known to play a role.

AHR mediated disorders are well established in the art. The nexusbetween AHR and AHR mediated disorders diseases and/or conditions asrecited herein is well established in the relevant arts. For example,see: Uyttenhove et al., “Evidence for a tumoral immune resistancemechanism based on tryptophan degradation by indoleamine2,3-dioxygenase” Nature Medicine, 2003 vol. 9(10), 1038; Murray et al.,“AH RECEPTOR LIGANDS IN CANCER: FRIEND AND FOE” Nat. Rev. CancerDecember 2014, vol. 14(12), pages 801-814; Moon et al., “Targeting theindoleamine 2,3-dioxygenase pathway in cancer” J. Immuno Therapy ofCancer, 2015 vol 3, page 51; Ishida et al., “Activation of arylhydrocarbon receptor promotes invasion of clear cell renal cellcarcinoma and is associated with poor prognosis and cigarette smoke”Int. J. Cancer July 2015 vol. 15, no. 137(2), pages 299-310; Ishida etal., “Activation of the aryl hydrocarbon receptor pathway enhancescancer cell invasion by upregulating the MMP expression and isassociated with poor prognosis in upper urinary tract urothelial cancer”Carcinogenesis February 2010 vol. 31(2), pages 287-295. Su et al.,“Prognostic value of nuclear translocation of aryl hydrocarbon receptorfor non-small cell lung cancer” Anticancer Res. September 2013, vol.33(9), pages 3953-3961; Peng et al., “Aryl hydrocarbon receptor pathwayactivation enhances gastric cancer cell invasiveness likely through ac-Jun-dependent induction of matrix metalloproteinase-9” BMC Cell Biol.April 2009 vol. 16; pages 10-27; Jin et al., “Aryl Hydrocarbon ReceptorActivation Reduces Dendritic Cell Function during Influenza VirusInfection” Toxicol Sci. August 2010, vol. 116(2), pages 514-522; Head etal., “The aryl hydrocarbon receptor is a modulator of anti-viralimmunity” Biochem. Pharmacol. February 2009 vol. 15; no. 77(4), pages642-53; Jin et al., “New insights into the role of the aryl hydrocarbonreceptor in the function of CD11c′ cells during respiratory viralinfection” Eur. J. Immunol. June 2014, vol. 44(6), pages 1685-98; Nguyenet al., “Aryl hydrocarbon receptor and kynurenine: recent advances inautoimmune disease research” Front Immunol. October 2014, vol. 29, no.5, page 551; Esser et al., “The aryl hydrocarbon receptor in immunity”Trends in Immunology, Vol. 30, No. 9.

In some embodiments, the present invention provides a method fortreating one or more disorders, diseases, and/or conditions wherein thedisorder, disease, or condition is a proliferative disease such ascancer, an inflammatory disorder, or a viral infection.

In certain embodiments, the present invention provides a method oftreating cancer or another proliferative disorder, comprisingadministering a compound or composition of the present invention to apatient with cancer or another proliferative disorder. In certainembodiments, the method of treating cancer or another proliferativedisorder comprises administering compounds and compositions of thepresent invention to a mammal. In certain embodiments, the mammal is ahuman.

As used herein, the terms “inhibition of cancer” and “inhibition ofcancer cell proliferation” refer to the inhibition of the growth,division, maturation or viability of cancer cells, and/or causing thedeath of cancer cells, individually or in aggregate with other cancercells, by cytotoxicity, nutrient depletion, or the induction ofapoptosis.

Examples of tissues containing cancerous cells whose proliferation isinhibited by the compounds and compositions described herein and againstwhich the methods described herein are useful include but are notlimited to breast, prostate, brain, blood, bone marrow, liver, pancreas,skin, kidney, colon, ovary, lung, testicle, penis, thyroid, parathyroid,pituitary, thymus, retina, uvea, conjunctiva, spleen, head, neck,trachea, gall bladder, rectum, salivary gland, adrenal gland, throat,esophagus, lymph nodes, sweat glands, sebaceous glands, muscle, heart,and stomach.

In some embodiments, the cancer treated by compounds or compositions ofthe invention is a melanoma, liposarcoma, lung cancer, breast cancer,prostate cancer, leukemia, kidney cancer, esophageal cancer, braincancer, lymphoma or colon cancer. In certain embodiments, the cancer isa primary effusion lymphoma (PEL).

Compounds of the current invention are useful in the treatment of aproliferative disease selected from a benign or malignant tumor,carcinoma of the brain, kidney, liver, adrenal gland, bladder, breast,stomach, gastric tumors, ovaries, colon, rectum, prostate, pancreas,lung, vagina, cervix, testis, genitourinary tract, esophagus, larynx,skin, bone or thyroid, sarcoma, glioblastomas, neuroblastomas, multiplemyeloma or gastrointestinal cancer, especially colon carcinoma orcolorectal adenoma or a tumor of the neck and head, an epidermalhyperproliferation, psoriasis, prostate hyperplasia, a neoplasia, aneoplasia of epithelial character, adenoma, adenocarcinoma,keratoacanthoma, epidermoid carcinoma, large cell carcinoma,non-small-cell lung carcinoma, lymphomas, Hodgkins and Non-Hodgkins,Waldenström's macroglobulinemia, a mammary carcinoma, follicularcarcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma,melanoma, an MYD88-driven disorder, DLBCL, ABC DLBCL, an IL-1-drivendisorder, Smoldering of indolent multiple myeloma, or a leukemia.

Cancer includes, in some embodiments, without limitation, leukemias(e.g., acute leukemia, acute lymphocytic leukemia, acute myelocyticleukemia, acute myeloblastic leukemia, acute promyelocytic leukemia,acute myelomonocytic leukemia, acute monocytic leukemia, acuteerythroleukemia, chronic leukemia, chronic myelocytic leukemia, chroniclymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin'sdisease or non-Hodgkin's disease), Waldenström's macroglobulinemia,multiple myeloma, heavy chain disease, and solid tumors such as sarcomasand carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma,chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma,endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma,synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma,rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer,ovarian cancer, prostate cancer, squamous cell carcinoma, basal cellcarcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous glandcarcinoma, papillary carcinoma, papillary adenocarcinomas,cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renalcell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma,seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterinecancer, testicular cancer, lung carcinoma, small cell lung carcinoma,bladder carcinoma, epithelial carcinoma, glioma, astrocytoma,glioblastoma multiforme (GBM, also known as glioblastoma),medulloblastoma, craniopharyngioma, ependymoma, pinealoma,hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma,neurofibrosarcoma, meningioma, melanoma, neuroblastoma, andretinoblastoma).

In some embodiments, the cancer is glioma, astrocytoma, glioblastomamultiforme (GBM, also known as glioblastoma), medulloblastoma,craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acousticneuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma,melanoma, neuroblastoma, or retinoblastoma.

In some embodiments, the cancer is acoustic neuroma, astrocytoma (e.g.Grade I Pilocytic Astrocytoma, Grade II Low-grade Astrocytoma, Grade IIIAnaplastic Astrocytoma, or Grade IV Glioblastoma (GBM)), chordoma, CNSlymphoma, craniopharyngioma, brain stem glioma, ependymoma, mixedglioma, optic nerve glioma, subependymoma, medulloblastoma, meningioma,metastatic brain tumor, oligodendroglioma, pituitary tumors, primitiveneuroectodermal (PNET) tumor, or schwannoma. In some embodiments, thecancer is a type found more commonly in children than adults, such asbrain stem glioma, craniopharyngioma, ependymoma, juvenile pilocyticastrocytoma (JPA), medulloblastoma, optic nerve glioma, pineal tumor,primitive neuroectodermal tumors (PNET), or rhabdoid tumor. In someembodiments, the patient is an adult human. In some embodiments, thepatient is a child or pediatric patient.

Cancer includes, in another embodiment, without limitation,mesothelioma, hepatobiliary (hepatic and biliary duct), bone cancer,pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous orintraocular melanoma, ovarian cancer, colon cancer, rectal cancer,cancer of the anal region, stomach cancer, gastrointestinal (gastric,colorectal, and duodenal), uterine cancer, carcinoma of the fallopiantubes, carcinoma of the endometrium, carcinoma of the cervix, carcinomaof the vagina, carcinoma of the vulva, Hodgkin's Disease, cancer of theesophagus, cancer of the small intestine, cancer of the endocrinesystem, cancer of the thyroid gland, cancer of the parathyroid gland,cancer of the adrenal gland, sarcoma of soft tissue, cancer of theurethra, cancer of the penis, prostate cancer, testicular cancer,chronic or acute leukemia, chronic myeloid leukemia, lymphocyticlymphomas, cancer of the bladder, cancer of the kidney or ureter, renalcell carcinoma, carcinoma of the renal pelvis, non-Hodgkins's lymphoma,spinal axis tumors, brain stem glioma, pituitary adenoma, adrenocorticalcancer, gall bladder cancer, multiple myeloma, cholangiocarcinoma,fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one ormore of the foregoing cancers.

In some embodiments, the cancer is selected from hepatocellularcarcinoma, ovarian cancer, ovarian epithelial cancer, or fallopian tubecancer; papillary serous cystadenocarcinoma or uterine papillary serouscarcinoma (UPSC); prostate cancer; testicular cancer; gallbladdercancer; hepatocholangiocarcinoma; soft tissue and bone synovial sarcoma;rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing sarcoma;anaplastic thyroid cancer; adrenocortical adenoma; pancreatic cancer;pancreatic ductal carcinoma or pancreatic adenocarcinoma;gastrointestinal/stomach (GIST) cancer; lymphoma; squamous cellcarcinoma of the head and neck (SCCHN); salivary gland cancer; glioma,or brain cancer; neurofibromatosis-1 associated malignant peripheralnerve sheath tumors (MPNST); Waldenstrom's macroglobulinemia; ormedulloblastoma.

In some embodiments, the cancer is selected from hepatocellularcarcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovariancancer, ovarian epithelial cancer, fallopian tube cancer, papillaryserous cystadenocarcinoma, uterine papillary serous carcinoma (UPSC),hepatocholangiocarcinoma, soft tissue and bone synovial sarcoma,rhabdomyosarcoma, osteosarcoma, anaplastic thyroid cancer,adrenocortical adenoma, pancreatic cancer, pancreatic ductal carcinoma,pancreatic adenocarcinoma, glioma, neurofibromatosis-1 associatedmalignant peripheral nerve sheath tumors (MPNST), Waldenstrom'smacroglobulinemia, or medulloblastoma.

In some embodiments, the cancer is a solid tumor, such as a sarcoma,carcinoma, or lymphoma. Solid tumors generally comprise an abnormal massof tissue that typically does not include cysts or liquid areas. In someembodiments, the cancer is selected from renal cell carcinoma, or kidneycancer; hepatocellular carcinoma (HCC) or hepatoblastoma, or livercancer; melanoma; breast cancer; colorectal carcinoma, or colorectalcancer; colon cancer; rectal cancer; anal cancer; lung cancer, such asnon-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC);ovarian cancer, ovarian epithelial cancer, ovarian carcinoma, orfallopian tube cancer; papillary serous cystadenocarcinoma or uterinepapillary serous carcinoma (UPSC); prostate cancer; testicular cancer;gallbladder cancer; hepatocholangiocarcinoma; soft tissue and bonesynovial sarcoma; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewingsarcoma; anaplastic thyroid cancer; adrenocortical carcinoma; pancreaticcancer; pancreatic ductal carcinoma or pancreatic adenocarcinoma;gastrointestinal/stomach (GIST) cancer; lymphoma; squamous cellcarcinoma of the head and neck (SCCHN); salivary gland cancer; glioma,or brain cancer; neurofibromatosis-1 associated malignant peripheralnerve sheath tumors (MPNST); Waldenstrom's macroglobulinemia; ormedulloblastoma.

In some embodiments, the cancer is selected from renal cell carcinoma,hepatocellular carcinoma (HCC), hepatoblastoma, colorectal carcinoma,colorectal cancer, colon cancer, rectal cancer, anal cancer, ovariancancer, ovarian epithelial cancer, ovarian carcinoma, fallopian tubecancer, papillary serous cystadenocarcinoma, uterine papillary serouscarcinoma (UPSC), hepatocholangiocarcinoma, soft tissue and bonesynovial sarcoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma,anaplastic thyroid cancer, adrenocortical carcinoma, pancreatic cancer,pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma, braincancer, neurofibromatosis-1 associated malignant peripheral nerve sheathtumors (MPNST), Waldenstrom's macroglobulinemia, or medulloblastoma.

In some embodiments, the cancer is selected from hepatocellularcarcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovariancancer, ovarian epithelial cancer, ovarian carcinoma, fallopian tubecancer, papillary serous cystadenocarcinoma, uterine papillary serouscarcinoma (UPSC), hepatocholangiocarcinoma, soft tissue and bonesynovial sarcoma, rhabdomyosarcoma, osteosarcoma, anaplastic thyroidcancer, adrenocortical carcinoma, pancreatic cancer, pancreatic ductalcarcinoma, pancreatic adenocarcinoma, glioma, neurofibromatosis-1associated malignant peripheral nerve sheath tumors (MPNST),Waldenstrom's macroglobulinemia, or medulloblastoma.

In some embodiments, the cancer is hepatocellular carcinoma (HCC). Insome embodiments, the cancer is hepatoblastoma. In some embodiments, thecancer is colon cancer. In some embodiments, the cancer is rectalcancer. In some embodiments, the cancer is ovarian cancer, or ovariancarcinoma. In some embodiments, the cancer is ovarian epithelial cancer.In some embodiments, the cancer is fallopian tube cancer. In someembodiments, the cancer is papillary serous cystadenocarcinoma. In someembodiments, the cancer is uterine papillary serous carcinoma (UPSC). Insome embodiments, the cancer is hepatocholangiocarcinoma. In someembodiments, the cancer is soft tissue and bone synovial sarcoma. Insome embodiments, the cancer is rhabdomyosarcoma. In some embodiments,the cancer is osteosarcoma. In some embodiments, the cancer isanaplastic thyroid cancer. In some embodiments, the cancer isadrenocortical carcinoma. In some embodiments, the cancer is pancreaticcancer, or pancreatic ductal carcinoma. In some embodiments, the canceris pancreatic adenocarcinoma. In some embodiments, the cancer is glioma.In some embodiments, the cancer is malignant peripheral nerve sheathtumors (MPNST). In some embodiments, the cancer is neurofibromatosis-1associated MPNST. In some embodiments, the cancer is Waldenstrom'smacroglobulinemia. In some embodiments, the cancer is medulloblastoma.

In some embodiments, the cancer is Acute Lymphoblastic Leukemia (ALL),Acute Myeloid Leukemia (AML), Adrenocortical Carcinoma, Anal Cancer,Appendix Cancer, Atypical Teratoid/Rhabdoid Tumor, Basal Cell Carcinoma,Bile Duct Cancer, Bladder Cancer, Bone Cancer, Brain Tumor, Astrocytoma,Brain and Spinal Cord Tumor, Brain Stem Glioma, Central Nervous SystemAtypical Teratoid/Rhabdoid Tumor, Central Nervous System EmbryonalTumors, Breast Cancer, Bronchial Tumors, Burkitt Lymphoma, CarcinoidTumor, Carcinoma of Unknown Primary, Central Nervous System Cancer,Cervical Cancer, Childhood Cancers, Chordoma, Chronic LymphocyticLeukemia (CLL), Chronic Myelogenous Leukemia (CML), ChronicMyeloproliferative Disorders, Colon Cancer, Colorectal Cancer,Craniopharyngioma, Cutaneous T-Cell Lymphoma, Ductal Carcinoma In Situ(DCIS), Embryonal Tumors, Endometrial Cancer, Ependymoblastoma,Ependymoma, Esophageal Cancer, Esthesioneuroblastoma, Ewing Sarcoma,Extracranial Germ Cell Tumor, Extragonadal Germ Cell Tumor, ExtrahepaticBile Duct Cancer, Eye Cancer, Fibrous Histiocytoma of Bone, GallbladderCancer, Gastric Cancer, Gastrointestinal Carcinoid Tumor,Gastrointestinal Stromal Tumors (GIST), Germ Cell Tumor, Ovarian GermCell Tumor, Gestational Trophoblastic Tumor, Glioma, Hairy CellLeukemia, Head and Neck Cancer, Heart Cancer, Hepatocellular Cancer,Histiocytosis, Langerhans Cell Cancer, Hodgkin Lymphoma, HypopharyngealCancer, Intraocular Melanoma, Islet Cell Tumors, Kaposi Sarcoma, KidneyCancer, Langerhans Cell Histiocytosis, Laryngeal Cancer, Leukemia, Lipand Oral Cavity Cancer, Liver Cancer, Lobular Carcinoma In Situ (LCIS),Lung Cancer, Lymphoma, AIDS-Related Lymphoma, Macroglobulinemia, MaleBreast Cancer, Medulloblastoma, Medulloepithelioma, Melanoma, MerkelCell Carcinoma, Malignant Mesothelioma, Metastatic Squamous Neck Cancerwith Occult Primary, Midline Tract Carcinoma Involving NUT Gene, MouthCancer, Multiple Endocrine Neoplasia Syndrome, Multiple Myeloma/PlasmaCell Neoplasm, Mycosis Fungoides, Myelodysplastic Syndrome,Myelodysplastic/Myeloproliferative Neoplasm, Chronic MyelogenousLeukemia (CML), Acute Myeloid Leukemia (AML), Myeloma, Multiple Myeloma,Chronic Myeloproliferative Disorder, Nasal Cavity Cancer, ParanasalSinus Cancer, Nasopharyngeal Cancer, Neuroblastoma, Non-HodgkinLymphoma, Non-Small Cell Lung Cancer, Oral Cancer, Oral Cavity Cancer,Lip Cancer, Oropharyngeal Cancer, Osteosarcoma, Ovarian Cancer,Pancreatic Cancer, Papillomatosis, Paraganglioma, Paranasal SinusCancer, Nasal Cavity Cancer, Parathyroid Cancer, Penile Cancer,Pharyngeal Cancer, Pheochromocytoma, Pineal Parenchymal Tumors ofIntermediate Differentiation, Pineoblastoma, Pituitary Tumor, PlasmaCell Neoplasm, Pleuropulmonary Blastoma, Breast Cancer, Primary CentralNervous System (CNS) Lymphoma, Prostate Cancer, Rectal Cancer, RenalCell Cancer, Clear cell renal cell carcinoma, Renal Pelvis Cancer,Ureter Cancer, Transitional Cell Cancer, Retinoblastoma,Rhabdomyosarcoma, Salivary Gland Cancer, Sarcoma, Sezary Syndrome, SkinCancer, Small Cell Lung Cancer, Small Intestine Cancer, Soft TissueSarcoma, Squamous Cell Carcinoma, Squamous Neck Cancer with OccultPrimary, Squamous Cell Carcinoma of the Head and Neck (HNSCC), StomachCancer, Supratentorial Primitive Neuroectodermal Tumors, T-CellLymphoma, Testicular Cancer, Throat Cancer, Thymoma, Thymic Carcinoma,Thyroid Cancer, Transitional Cell Cancer of the Renal Pelvis and Ureter,Triple Negative Breast Cancer (TNBC), Gestational Trophoblastic Tumor,Unknown Primary, Unusual Cancer of Childhood, Urethral Cancer, UterineCancer, Uterine Sarcoma, Waldenstrom Macroglobulinemia, or Wilms Tumor.

Compounds according to the invention are useful in the treatment ofinflammatory or obstructive airways diseases, resulting, for example, inreduction of tissue damage, airways inflammation, bronchialhyperreactivity, remodeling or disease progression. Inflammatory orobstructive airways diseases to which the present invention isapplicable include asthma of whatever type or genesis including bothintrinsic (non-allergic) asthma and extrinsic (allergic) asthma, mildasthma, moderate asthma, severe asthma, bronchitic asthma,exercise-induced asthma, occupational asthma and asthma inducedfollowing bacterial infection. Treatment of asthma is also to beunderstood as embracing treatment of subjects, e.g. of less than 4 or 5years of age, exhibiting wheezing symptoms and diagnosed or diagnosableas “wheezy infants”, an established patient category of major medicalconcern and now often identified as incipient or early-phase asthmatics.

Prophylactic efficacy in the treatment of asthma will be evidenced byreduced frequency or severity of symptomatic attack, e.g. of acuteasthmatic or bronchoconstrictor attack, improvement in lung function orimproved airways hyperreactivity. It may further be evidenced by reducedrequirement for other, symptomatic therapy, such as therapy for orintended to restrict or abort symptomatic attack when it occurs, forexample antiinflammatory or bronchodilatory. Prophylactic benefit inasthma may in particular be apparent in subjects prone to “morningdipping”. “Morning dipping” is a recognized asthmatic syndrome, commonto a substantial percentage of asthmatics and characterised by asthmaattack, e.g. between the hours of about 4 to 6 am, i.e. at a timenormally substantially distant form any previously administeredsymptomatic asthma therapy.

Compounds of the current invention can be used for other inflammatory orobstructive airways diseases and conditions to which the presentinvention is applicable and include acute lung injury (ALI), adult/acuterespiratory distress syndrome (ARDS), chronic obstructive pulmonary,airways or lung disease (COPD, COAD or COLD), including chronicbronchitis or dyspnea associated therewith, emphysema, as well asexacerbation of airways hyperreactivity consequent to other drugtherapy, in particular other inhaled drug therapy. The invention is alsoapplicable to the treatment of bronchitis of whatever type or genesisincluding, but not limited to, acute, arachidic, catarrhal, croupus,chronic or phthinoid bronchitis. Further inflammatory or obstructiveairways diseases to which the present invention is applicable includepneumoconiosis (an inflammatory, commonly occupational, disease of thelungs, frequently accompanied by airways obstruction, whether chronic oracute, and occasioned by repeated inhalation of dusts) of whatever typeor genesis, including, for example, aluminosis, anthracosis, asbestosis,chalicosis, ptilosis, siderosis, silicosis, tabacosis and byssinosis.

With regard to their anti-inflammatory activity, in particular inrelation to inhibition of eosinophil activation, compounds of theinvention are also useful in the treatment of eosinophil relateddisorders, e.g. eosinophilia, in particular eosinophil related disordersof the airways (e.g. involving morbid eosinophilic infiltration ofpulmonary tissues) including hypereosinophilia as it effects the airwaysand/or lungs as well as, for example, eosinophil-related disorders ofthe airways consequential or concomitant to Loffler's syndrome,eosinophilic pneumonia, parasitic (in particular metazoan) infestation(including tropical eosinophilia), bronchopulmonary aspergillosis,polyarteritis nodosa (including Churg-Strauss syndrome), eosinophilicgranuloma and eosinophil-related disorders affecting the airwaysoccasioned by drug-reaction.

Compounds of the invention are also useful in the treatment ofinflammatory or allergic conditions of the skin, for example psoriasis,contact dermatitis, atopic dermatitis, alopecia areata, erythemamultiforma, dermatitis herpetiformis, scleroderma, vitiligo,hypersensitivity angiitis, urticaria, bullous pemphigoid, lupuserythematosus, systemic lupus erythematosus, pemphigus vulgaris,pemphigus foliaceus, paraneoplastic pemphigus, epidermolysis bullosaacquisita, acne vulgaris, and other inflammatory or allergic conditionsof the skin.

Compounds of the invention may also be used for the treatment of otherdiseases or conditions, such as diseases or conditions having aninflammatory component, for example, treatment of diseases andconditions of the eye such as ocular allergy, conjunctivitis,keratoconjunctivitis sicca, and vernal conjunctivitis, diseasesaffecting the nose including allergic rhinitis, and inflammatory diseasein which autoimmune reactions are implicated or having an autoimmunecomponent or etiology, including autoimmune hematological disorders(e.g. hemolytic anemia, aplastic anemia, pure red cell anemia andidiopathic thrombocytopenia), systemic lupus erythematosus, rheumatoidarthritis, polychondritis, scleroderma, Wegener granulamatosis,dermatomyositis, chronic active hepatitis, myasthenia gravis,Steven-Johnson syndrome, idiopathic sprue, autoimmune inflammatory boweldisease (e.g. ulcerative colitis and Crohn's disease), irritable bowelsyndrome, celiac disease, periodontitis, hyaline membrane disease,kidney disease, glomerular disease, alcoholic liver disease, multiplesclerosis, endocrine opthalmopathy, Graves' disease, sarcoidosis,alveolitis, chronic hypersensitivity pneumonitis, multiple sclerosis,primary biliary cirrhosis, uveitis (anterior and posterior), Sjogren'ssyndrome, keratoconjunctivitis sicca and vernal keratoconjunctivitis,interstitial lung fibrosis, psoriatic arthritis, systemic juvenileidiopathic arthritis, cryopyrin-associated periodic syndrome, nephritis,vasculitis, diverticulitis, interstitial cystitis, glomerulonephritis(with and without nephrotic syndrome, e.g. including idiopathicnephrotic syndrome or minal change nephropathy), chronic granulomatousdisease, endometriosis, leptospiriosis renal disease, glaucoma, retinaldisease, ageing, headache, pain, complex regional pain syndrome, cardiachypertrophy, musclewasting, catabolic disorders, obesity, fetal growthretardation, hyperchlolesterolemia, heart disease, chronic heartfailure, mesothelioma, anhidrotic ecodermal dysplasia, Behcet's disease,incontinentia pigmenti, Paget's disease, pancreatitis, hereditaryperiodic fever syndrome, asthma (allergic and non-allergic, mild,moderate, severe, bronchitic, and exercise-induced), acute lung injury,acute respiratory distress syndrome, eosinophilia, hypersensitivities,anaphylaxis, nasal sinusitis, ocular allergy, silica induced diseases,COPD (reduction of damage, airways inflammation, bronchialhyperreactivity, remodeling or disease progression), pulmonary disease,cystic fibrosis, acid-induced lung injury, pulmonary hypertension,polyneuropathy, cataracts, muscle inflammation in conjunction withsystemic sclerosis, dermatomyositis, polymyositis, inclusion bodymyositis, myasthenia gravis, thyroiditis, Addison's disease, lichenplanus, Type 1 diabetes, or Type 2 diabetes.

In some embodiments the inflammatory disease which can be treatedaccording to the methods of this invention is selected from acute andchronic gout, chronic gouty arthritis, psoriasis, psoriatic arthritis,rheumatoid arthritis, Juvenile rheumatoid arthritis, Systemic juvenileidiopathic arthritis (SJIA), Cryopyrin-Associated Periodic Syndromes(CAPS), or osteoarthritis.

In some embodiments, the inflammatory disease which can be treatedaccording to the methods of this invention is selected from aTH17-mediated disease. In some embodiments, the TH17-mediated disease isselected from Systemic lupus erythematosus, Multiple sclerosis,inflammatory bowel disease including Crohn's or ulcerative colitis.

In some embodiments, the inflammatory disease which can be treatedaccording to the methods of this invention is selected from Sjogren'ssyndrome allergic disorders, osteoarthritis. Conditions of the eye suchas ocular allergy, conjunctivitis, keratoconjunctivitis sicca, andvernal conjunctivitis, diseases affecting the nose including allergicrhinitis.

In some embodiments, the inflammatory disease which can be treatedaccording to the methods of this invention is selected from contactdermatitis, atopic dermatitis, alopecia areata, erythema multiforma,dermatitis herpetiformis, scleroderma, vitiligo, hypersensitivityangiitis, urticaria, bullous pemphigoid, pemphigus vulgaris, pemphigusfoliaceus, paraneoplastic pemphigus, epidermolysis bullosa acquisita,and other inflammatory or allergic conditions of the skin.

In certain embodiments, a provided compound is useful for treating aviral infection, disease, or condition. In some embodiments, the presentinvention provides a method of treating a viral disease selected fromretroviral diseases, such as, HIV-1, HIV-2, human T-cell leukemiavirus-I (HTLV-I), HTLV-II, HTLV-III, simian immunodeficiency virus(SIV), lymphadenopathy-associated virus (LAV-2), simian T-lymphotrophicvirus-I (STLV-I), STLV-II, STLV-III, simian B-lymphotrophic (SBL) virus,Gibbon ape leukemia virus (GALV), bovine leukemia virus (BLV), equineinfectious anemia virus (EIAV), feline leukemia virus (FELV), murineleukemia virus (MuLV), avian leukosis virus (ALV); other virusinfections such as hepadnaviridae (Hepatitis B); herpesviridae (Herpessimplex I, Herpes simplex II, Varicella-Zoster, Epstein-Barr virus andcytomegalovirus); parvoviridae (human parvovirus B-19); papovaviridae(human papilloma virus types 1 to 60, JC and BK viruses); pox viruses(variola major, variola minor, vaccinia, monkey pox, cowpox,paravaccinia or milker's node virus, parapox or ORF virus, molluscumcontagiosum) and cancers, lymphomas and other leukemias.

Combination Therapies

Depending upon the particular condition, or disease, to be treated,additional therapeutic agents, which are normally administered to treatthat condition, may be administered in combination with compounds andcompositions of this invention. As used herein, additional therapeuticagents that are normally administered to treat a particular disease, orcondition, are known as “appropriate for the disease, or condition,being treated”.

In certain embodiments, a provided compound, or a composition thereof,is administered in combination with another anti-cancer, cytotoxin, orchemotherapeutic agent, to a patient in need thereof.

In certain embodiments, the anti-cancer or chemotherapeutic agents usedin combination with compounds or compositions of the invention include,but are not limited to metformin, phenformin, buformin, imatinib,nilotinib, gefitinib, sunitinib, carfilzomib, salinosporamide A,retinoic acid, cisplatin, carboplatin, oxaliplatin, mechlorethamine,cyclophosphamide, chlorambucil, ifosfamide, azathioprine,mercaptopurine, doxifluridine, fluorouracil, gemcitabine, methotrexate,tioguanine, vincristine, vinblastine, vinorelbine, vindesine,podophyllotoxin, etoposide, teniposide, tafluposide, paclitaxel,docetaxel, irinotecan, topotecan, amsacrine, actinomycin, doxorubicin,daunorubicin, valrubicin, idarubicin, epirubicin, plicamycin, mitomycin,mitoxantrone, melphalan, busulfan, capecitabine, pemetrexed,epothilones, 13-cis-Retinoic Acid, 2-CdA, 2-Chlorodeoxyadenosine,5-Azacitidine, 5-Fluorouracil, 5-FU, 6-Mercaptopurine, 6-MP, 6-TG,6-Thioguanine, Abraxane, Accutane®, Actinomycin-D, Adriamycin®,Adrucil®, Afinitor®, Agrylin®, Ala-Cort®, Aldesleukin, Alemtuzumab,ALIMTA, Alitretinoin, Alkaban-AQ⁻, Alkeran®, All-transretinoic Acid,Alpha Interferon, Altretamine, Amethopterin, Amifostine,Aminoglutethimide, Anagrelide, Anandron®, Anastrozole,Arabinosylcytosine, Ara-C, Aranesp®, Aredia®, Arimidex®, Aromasin®,Arranon®, Arsenic Trioxide, Arzerra™, Asparaginase, ATRA, Avastin®,Azacitidine, BCG, BCNU, Bendamustine, Bevacizumab, Bexarotene, BEXXAR®,Bicalutamide, BiCNU, Blenoxane®, Bleomycin, Bortezomib, Busulfan,Busulfex®, C225, Calcium Leucovorin, Campath®, Camptosar®,Camptothecin-11, Capecitabine, Carac™, Carboplatin, Carmustine,Carmustine Wafer, Casodex®, CC-5013, CCI-779, CCNU, CDDP, CeeNU,Cerubidine®, Cetuximab, Chlorambucil, Citrovorum Factor, Cladribine,Cortisone, Cosmegen®, CPT-11, Cytadren®, Cytosar-U®, Cytoxan®,Dacarbazine, Dacogen, Dactinomycin, Darbepoetin Alfa, Dasatinib,Daunomycin, Daunorubicin Hydrochloride, Daunorubicin Liposomal,DaunoXome®, Decadron, Decitabine, Delta-Cortef®, Deltasone®, Denileukin,Diftitox, DepoCyt™, Dexamethasone, Dexamethasone Acetate, DexamethasoneSodium Phosphate, Dexasone, Dexrazoxane, DHAD, DIC, Diodex, Docetaxel,Doxil®, Doxorubicin, Doxorubicin Liposomal, Droxia™, DTIC, DTIC-Dome®,Duralone®, Efudex®, Eligard™, Ellence™, Eloxatin™, Elspar®, Emcyt®,Epirubicin, Epoetin Alfa, Erbitux, Erlotinib, Erwinia L-asparaginase,Estramustine, Ethyol, Etopophos®, Etoposide, Etoposide Phosphate,Eulexin®, Everolimus, Evista®, Exemestane, Fareston®, Faslodex®,Femara®, Filgrastim, Floxuridine, Fludara®, Fludarabine, Fluoroplex®,Fluorouracil, Fluorouracil (cream), Fluoxymesterone, Flutamide, FolinicAcid, FUDR®, Fulvestrant, G-CSF, Gefitinib, Gemcitabine, Gemtuzumab,ozogamicin, Gemzar Gleevec™ Gliadel® Wafer, GM-CSF, Goserelin,Granulocyte—Colony Stimulating Factor, Granulocyte Macrophage ColonyStimulating Factor, Halotestin®, Herceptin®, Hexadrol, Hexalen®,Hexamethylmelamine, HMM, Hycamtin®, Hydrea®, Hydrocort Acetate®,Hydrocortisone, Hydrocortisone Sodium Phosphate, Hydrocortisone SodiumSuccinate, Hydrocortone Phosphate, Hydroxyurea, Ibritumomab,Ibritumomab, Tiuxetan, Idamycin®, Idarubicin Ifex®, IFN-alpha,Ifosfamide, IL-11, IL-2, Imatinib mesylate, Imidazole Carboxamide,Interferon alfa, Interferon Alfa-2b (PEG Conjugate), Interleukin-2,Interleukin-11, Intron A® (interferon alfa-2b), Iressa®, Irinotecan,Isotretinoin, Ixabepilone, Ixempra™, Kidrolase®, Lanacort®, Lapatinib,L-asparaginase, LCR, Lenalidomide, Letrozole, Leucovorin, Leukeran,Leukine™, Leuprolide, Leurocristine, Leustatin™, Liposomal Ara-C, LiquidPred®, Lomustine, L-PAM, L-Sarcolysin, Lupron®, Lupron Depot®,Matulane®, Maxidex, Mechlorethamine, Mechlorethamine Hydrochloride,Medralone®, Medrol®, Megace®, Megestrol, Megestrol Acetate, Melphalan,Mercaptopurine, Mesna, Mesnex™, Methotrexate, Methotrexate Sodium,Methylprednisolone, Meticorten®, Mitomycin, Mitomycin-C, Mitoxantrone,M-Prednisol®, MTC, MTX, Mustargen®, Mustine, Mutamycin®, Myleran®,Mylocel™, Mylotarg®, Navelbine®, Nelarabine, Neosar®, Neulasta™,Neumega®, Neupogen®, Nexavar®, Nilandron®, Nilotinib, Nilutamide,Nipent®, Nitrogen Mustard, Novaldex®, Novantrone®, Nplate, Octreotide,Octreotide acetate, Ofatumumab, Oncospar®, Oncovin®, Ontak®, Onxal™,Oprelvekin, Orapred®, Orasone®, Oxaliplatin, Paclitaxel, PaclitaxelProtein-bound, Pamidronate, Panitumumab, Panretin®, Paraplatin®,Pazopanib, Pediapred®, PEG Interferon, Pegaspargase, Pegfilgrastim,PEG-INTRON™, PEG-L-asparaginase, PEMETREXED, Pentostatin, PhenylalanineMustard, Platinol®, Platinol-AQ®, Prednisolone, Prednisone, Prelone®,Procarbazine, PROCRIT®, Proleukin®, Prolifeprospan 20 with CarmustineImplant, Purinethol®, Raloxifene, Revlimid®, Rheumatrex®, Rituxan®,Rituximab, Roferon-A® (Interferon Alfa-2a), Romiplostim, Rubex®,Rubidomycin hydrochloride, Sandostatin®, Sandostatin LAR®, Sargramostim,Solu-Cortef®, Solu-Medrol®, Sorafenib, SPRYCEL™, STI-571, Streptozocin,SU11248, Sunitinib, Sutent®, Tamoxifen, Tarceva®, Targretin®, Tasigna®,Taxol®, Taxotere®, Temodar®, Temozolomide, Temsirolimus, Teniposide,TESPA, Thalidomide, Thalomid®, TheraCys®, Thioguanine, ThioguanineTabloid®, Thiophosphoamide, Thioplex®, Thiotepa, TICE®, Toposar®,Topotecan, Toremifene, Torisel®, Tositumomab, Trastuzumab, Treanda®,Tretinoin, Trexall™ Trisenox®, TSPA, TYKERB®, VCR, Vectibix™, Velban®,Velcade®, VePesid®, Vesanoid®, Viadur™, Vidaza®, Vinblastine,Vinblastine Sulfate, Vincasar Pfs®, Vincristine, Vinorelbine,Vinorelbine tartrate, VLB, VM-26, Vorinostat, Votrient, VP-16, Vumon,Xeloda®, Zanosar®, Zevalin™, Zinecard®, Zoladex®, Zoledronic acid,Zolinza, Zometa®, or combinations of any of the above.

In certain embodiments, an immuno-oncology agent can be administeredwith a compound as described herein for treatment of a proliferativedisorder as described herein. As used herein, the term “animmuno-oncology agent” refers to an agent which is effective to enhance,stimulate, and/or up-regulate immune responses in a subject. In someembodiments, the administration of an immuno-oncology agent with acompound as described herein has a synergic effect in treating cancer.

In some embodiments, a compound as described herein is sequentiallyadministered prior to administration of an immuno-oncology agent. Insome embodiments, a compound as described herein is administeredconcurrently with an immuno-oncology agent. In some embodiments, acompound as described herein is sequentially administered afteradministration of an immuno-oncology agent.

In some embodiments, a compound as described herein may be co-formulatedwith an immuno-oncology agent.

An immuno-oncology agent can be, for example, a small molecule drug, anantibody, or a biologic or small molecule. Examples of biologicimmuno-oncology agents include, but are not limited to, cancer vaccines,antibodies, and cytokines. In some embodiments, an antibody is amonoclonal antibody. In some embodiments, a monoclonal antibody ishumanized or human.

In some embodiments, an immuno-oncology agent is (i) an agonist of astimulatory (including a co-stimulatory) receptor or (ii) an antagonistof an inhibitory (including a co-inhibitory) signal on T cells, both ofwhich result in amplifying antigen-specific T cell responses.

Certain of the stimulatory and inhibitory molecules are members of theimmunoglobulin super family (IgSF). One important family ofmembrane-bound ligands that bind to co-stimulatory or co-inhibitoryreceptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1),B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6.Another family of membrane bound ligands that bind to co-stimulatory orco-inhibitory receptors is the TNF family of molecules that bind tocognate TNF receptor family members, which includes CD40 and CD40L,OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1B),TRAIL/Apo2-L, TRAILR1/DR4, TRAILR2/DR5, TRAILR3, TRAILR4, OPG, RANK,RANKL, TWEAKR/Fn14, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LTβR,LIGHT, DcR3, HVEM, VEGI/TL1A, TRAMP/DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1,Lymphotoxin α/TNFβ, TNFR2, TNFα, LTβR, Lymphotoxin α1β2, FAS, FASL,RELT, DR6, TROY, NGFR.

In some embodiments, an immuno-oncology agent is a cytokine thatinhibits T cell activation (e.g., IL-6, IL-10, TGF-β, VEGF, and otherimmunosuppressive cytokines) or a cytokine that stimulates T cellactivation, for stimulating an immune response.

In some embodiments, a combination of a compound as described herein,and an immuno-oncology agent can stimulate T cell responses. In someembodiments, an immuno-oncology agent is: (i) an antagonist of a proteinthat inhibits T cell activation (e.g., immune checkpoint inhibitors)such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, Galectin 9, CEACAM-1,BTLA, CD69, Galectin-1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP,PD1H, LAIR1, TIM-1, and TTM-4; or (ii) an agonist of a protein thatstimulates T cell activation such as B7-1, B7-2, CD28, 4-1BB (CD137),4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3and CD28H.

In some embodiments, an immuno-oncology agent is an antagonist ofinhibitory receptors on NK cells or an agonists of activating receptorson NK cells. In some embodiments, an immuno-oncology agent is anantagonists of KIR, such as lirilumab.

In some embodiments, an immuno-oncology agent is an agent that inhibitsor depletes macrophages or monocytes, including but not limited toCSF-1R antagonists such as CSF-1R antagonist antibodies including RG7155(WO11/70024, WO11/107553, WO11/131407, WO13/87699, WO13/119716,WO13/132044) or FPA-008 (WO11/140249; WO13169264; WO14/036357).

In some embodiments, an immuno-oncology agent is selected from agonisticagents that ligate positive costimulatory receptors, blocking agentsthat attenuate signaling through inhibitory receptors, antagonists, andone or more agents that increase systemically the frequency ofanti-tumor T cells, agents that overcome distinct immune suppressivepathways within the tumor microenvironment (e.g., block inhibitoryreceptor engagement (e.g., PD-L1/PD-1 interactions), deplete or inhibitTregs (e.g., using an anti-CD25 monoclonal antibody (e.g., daclizumab)or by ex vivo anti-CD25 bead depletion), inhibit metabolic enzymes suchas IDO, or reverse/prevent T cell energy or exhaustion) and agents thattrigger innate immune activation and/or inflammation at tumor sites.

In some embodiments, an immuno-oncology agent is a CTLA-4 antagonist. Insome embodiments, a CTLA-4 antagonist is an antagonistic CTLA-4antibody. In some embodiments, an antagonistic CTLA-4 antibody is YERVOY(ipilimumab) or tremelimumab.

In some embodiments, an immuno-oncology agent is a PD-1 antagonist. Insome embodiments, a PD-1 antagonist is administered by infusion. In someembodiments, an immuno-oncology agent is an antibody or anantigen-binding portion thereof that binds specifically to a ProgrammedDeath-1 (PD-1) receptor and inhibits PD-1 activity. In some embodiments,a PD-1 antagonist is an antagonistic PD-1 antibody. In some embodiments,an antagonistic PD-1 antibody is OPDIVO (nivolumab), KEYTRUDA(pembrolizumab), or MEDI-0680 (AMP-514; WO2012/145493). In someembodiments, an immuno-oncology agent may be pidilizumab (CT-011). Insome embodiments, an immuno-oncology agent is a recombinant proteincomposed of the extracellular domain of PD-L2 (B7-DC) fused to the Fcportion of IgG, called AMP-224.

In some embodiments, an immuno-oncology agent is a PD-L1 antagonist. Insome embodiments, a PD-L1 antagonist is an antagonistic PD-L1 antibody.In some embodiments, a PD-L1 antibody is MPDL3280A (RG7446;WO2010/077634), durvalumab (MEDI4736), BMS-936559 (WO2007/005874), andMSB0010718C (WO2013/79174).

In some embodiments, an immuno-oncology agent is a LAG-3 antagonist. Insome embodiments, a LAG-3 antagonist is an antagonistic LAG-3 antibody.In some embodiments, a LAG3 antibody is BMS-986016 (WO10/19570,WO14/08218), or IMP-731 or MP-321 (WO08/132601, WO009/44273).

In some embodiments, an immuno-oncology agent is a CD137 (4-1B) agonist.In some embodiments, a CD137 (4-1BB) agonist is an agonistic CD137antibody. In some embodiments, a CD137 antibody is urelumab orPF-05082566 (WO12/32433).

In some embodiments, an immuno-oncology agent is a GITR agonist. In someembodiments, a GITR agonist is an agonistic GITR antibody. In someembodiments, a GITR antibody is BMS-986153, BMS-986156, TRX-518(WO006/105021, WO009/009116), or MK-4166 (WO11/028683).

In some embodiments, an immuno-oncology agent is an IDO antagonist. Insome embodiments, an IDO antagonist is INCB-024360 (WO2006/122150,WO07/75598, WO08/36653, WO08/36642), indoximod, or NLG-919 (WO09/73620,WO009/1156652, WO11/56652, WO12/142237).

In some embodiments, an immuno-oncology agent is an OX40 agonist. Insome embodiments, an OX40 agonist is an agonistic OX40 antibody. In someembodiments, an OX40 antibody is MEDI-6383 or MEDI-6469.

In some embodiments, an immuno-oncology agent is an OX40L antagonist. Insome embodiments, an OX40L antagonist is an antagonistic OX40 antibody.In some embodiments, an OX40L antagonist is RG-7888 (WO06/029879).

In some embodiments, an immuno-oncology agent is a CD40 agonist. In someembodiments, a CD40 agonist is an agonistic CD40 antibody. In someembodiments, an immuno-oncology agent is a CD40 antagonist. In someembodiments, a CD40 antagonist is an antagonistic CD40 antibody. In someembodiments, a CD40 antibody is lucatumumab or dacetuzumab.

In some embodiments, an immuno-oncology agent is a CD27 agonist. In someembodiments, a CD27 agonist is an agonistic CD27 antibody. In someembodiments, a CD27 antibody is varlilumab.

In some embodiments, an immuno-oncology agent is MGA271 (to B7H3)(WO11/109400).

In some embodiments, an immuno-oncology agent is abagovomab,adecatumumab, afutuzumab, alemtuzumab, anatumomab mafenatox, apolizumab,atezolimab, avelumab, blinatumomab, BMS-936559, catumaxomab, durvalumab,epacadostat, epratuzumab, indoximod, inotuzumab ozogamicin, intelumumab,ipilimumab, isatuximab, lambrolizumab, MED14736, MPDL3280A, nivolumab,obinutuzumab, ocaratuzumab, ofatumumab, olatatumab, pembrolizumab,pidilizumab, rituximab, ticilimumab, samalizumab, or tremelimumab.

In some embodiments, an immuno-oncology agent is an immunostimulatoryagent. For example, antibodies blocking the PD-1 and PD-L1 inhibitoryaxis can unleash activated tumor-reactive T cells and have been shown inclinical trials to induce durable anti-tumor responses in increasingnumbers of tumor histologies, including some tumor types thatconventionally have not been considered immunotherapy sensitive. See,e.g., Okazaki, T. et al. (2013) Nat. Immunol. 14, 1212-1218; Zou et al.(2016) Sci. Transl. Med. 8. The anti-PD-1 antibody nivolumab (Opdivo®,Bristol-Myers Squibb, also known as ONO-4538, MDX1106 and BMS-936558),has shown potential to improve the overall survival in patients with RCCwho had experienced disease progression during or after prioranti-angiogenic therapy.

In some embodiments, the immunomodulatory therapeutic specificallyinduces apoptosis of tumor cells. Approved immunomodulatory therapeuticswhich may be used in the present invention include pomalidomide(Pomalyst®, Celgene); lenalidomide (Revlimid®, Celgene); ingenolmebutate (Picato®, LEO Pharma).

In some embodiments, an immuno-oncology agent is a cancer vaccine. Insome embodiments, the cancer vaccine is selected from sipuleucel-T(Provenge®, Dendreon/Valeant Pharmaceuticals), which has been approvedfor treatment of asymptomatic, or minimally symptomatic metastaticcastrate-resistant (hormone-refractory) prostate cancer; and talimogenelaherparepvec (Imlygic®, BioVex/Amgen, previously known as T-VEC), agenetically modified oncolytic viral therapy approved for treatment ofunresectable cutaneous, subcutaneous and nodal lesions in melanoma. Insome embodiments, an immuno-oncology agent is selected from an oncolyticviral therapy such as pexastimogene devacirepvec (PexaVec/JX-594,SillaJen/formerly Jennerex Biotherapeutics), a thymidine kinase- (TK-)deficient vaccinia virus engineered to express GM-CSF, forhepatocellular carcinoma (NCT02562755) and melanoma (NCT00429312);pelareorep (Reolysin®, Oncolytics Biotech), a variant of respiratoryenteric orphan virus (reovirus) which does not replicate in cells thatare not RAS-activated, in numerous cancers, including colorectal cancer(NCT01622543); prostate cancer (NCT01619813); head and neck squamouscell cancer (NCT01166542); pancreatic adenocarcinoma (NCT00998322); andnon-small cell lung cancer (NSCLC) (NCT 00861627); enadenotucirev(NG-348, PsiOxus, formerly known as ColoAd1), an adenovirus engineeredto express a full length CD80 and an antibody fragment specific for theT-cell receptor CD3 protein, in ovarian cancer (NCT02028117); metastaticor advanced epithelial tumors such as in colorectal cancer, bladdercancer, head and neck squamous cell carcinoma and salivary gland cancer(NCT02636036); ONCOS-102 (Targovax/formerly Oncos), an adenovirusengineered to express GM-CSF, in melanoma (NCT03003676); and peritonealdisease, colorectal cancer or ovarian cancer (NCT02963831); GL-ONC1(GLV-1h68/GLV-1h153, Genelux GmbH), vaccinia viruses engineered toexpress beta-galactosidase (beta-gal)/beta-glucoronidase orbeta-gal/human sodium iodide symporter (hNIS), respectively, werestudied in peritoneal carcinomatosis (NCT01443260); fallopian tubecancer, ovarian cancer (NCT 02759588); or CG0070 (Cold Genesys), anadenovirus engineered to express GM-CSF, in bladder cancer(NCT02365818).

In some embodiments, an immuno-oncology agent is selected from JX-929(SillaJen/formerly Jennerex Biotherapeutics), a TK- and vaccinia growthfactor-deficient vaccinia virus engineered to express cytosinedeaminase, which is able to convert the prodrug 5-fluorocytosine to thecytotoxic drug 5-fluorouracil; TG01 and TG02 (Targovax/formerly Oncos),peptide-based immunotherapy agents targeted for difficult-to-treat RASmutations; and TILT-123 (TILT Biotherapeutics), an engineered adenovirusdesignated: Ad5/3-E2F-delta24-hTNFα-IRES-hIL20; and VSV-GP(ViraTherapeutics) a vesicular stomatitis virus (VSV) engineered toexpress the glycoprotein (GP) of lymphocytic choriomeningitis virus(LCMV), which can be further engineered to express antigens designed toraise an antigen-specific CD8⁺ T cell response.

In some embodiments, an immuno-oncology agent is a T-cell engineered toexpress a chimeric antigen receptor, or CAR. The T-cells engineered toexpress such chimeric antigen receptor are referred to as a CAR-T cells.

CARs have been constructed that consist of binding domains, which may bederived from natural ligands, single chain variable fragments (scFv)derived from monoclonal antibodies specific for cell-surface antigens,fused to endodomains that are the functional end of the T-cell receptor(TCR), such as the CD3-zeta signaling domain from TCRs, which is capableof generating an activation signal in T lymphocytes. Upon antigenbinding, such CARs link to endogenous signaling pathways in the effectorcell and generate activating signals similar to those initiated by theTCR complex.

For example, in some embodiments the CAR-T cell is one of thosedescribed in U.S. Pat. No. 8,906,682 (June; hereby incorporated byreference in its entirety), which discloses CAR-T cells engineered tocomprise an extracellular domain having an antigen binding domain (suchas a domain that binds to CD19), fused to an intracellular signalingdomain of the T cell antigen receptor complex zeta chain (such as CD3zeta). When expressed in the T cell, the CAR is able to redirect antigenrecognition based on the antigen binding specificity. In the case ofCD19, the antigen is expressed on malignant B cells. Over 200 clinicaltrials are currently in progress employing CAR-T in a wide range ofindications.[https://clinicaltrials.gov/ct2/results?term=chimeric+antigen+receptors&pg=1].

In some embodiments, an immunostimulatory agent is an activator ofretinoic acid receptor-related orphan receptor γ (RORγt). RORγt is atranscription factor with key roles in the differentiation andmaintenance of Type 17 effector subsets of CD4+ (Th17) and CD8+ (Tc17) Tcells, as well as the differentiation of IL-17 expressing innate immunecell subpopulations such as NK cells. In some embodiments, an activatorof RORγt is LYC-55716 (Lycera), which is currently being evaluated inclinical trials for the treatment of solid tumors (NCT02929862).

In some embodiments, an immunostimulatory agent is an agonist oractivator of a toll-like receptor (TLR). Suitable activators of TLRsinclude an agonist or activator of TLR9 such as SD-101 (Dynavax). SD-101is an immunostimulatory CpG which is being studied for B-cell,follicular and other lymphomas (NCT02254772). Agonists or activators ofTLR8 which may be used in the present invention include motolimod(VTX-2337, VentiRx Pharmaceuticals) which is being studied for squamouscell cancer of the head and neck (NCT02124850) and ovarian cancer(NCT02431559).

Other immuno-oncology agents that may be used in the present inventioninclude urelumab (BMS-663513, Bristol-Myers Squibb), an anti-CD137monoclonal antibody; varlilumab (CDX-1127, Celldex Therapeutics), ananti-CD27 monoclonal antibody; BMS-986178 (Bristol-Myers Squibb), ananti-OX40 monoclonal antibody; lirilumab (IPH2102/BMS-986015, InnatePharma, Bristol-Myers Squibb), an anti-KIR monoclonal antibody;monalizumab (IPH2201, Innate Pharma, AstraZeneca) an anti-NKG2Amonoclonal antibody; andecaliximab (GS-5745, Gilead Sciences), ananti-MMP9 antibody; MK-4166 (Merck & Co.), an anti-GITR monoclonalantibody.

In some embodiments, an immunostimulatory agent is selected fromelotuzumab, mifamurtide, an agonist or activator of a toll-likereceptor, and an activator of RORγt.

In some embodiments, an immunostimulatory therapeutic is recombinanthuman interleukin 15 (rhIL-15). rhIL-15 has been tested in the clinic asa therapy for melanoma and renal cell carcinoma (NCT01021059 andNCT01369888) and leukemias (NCT02689453). In some embodiments, animmunostimulatory agent is recombinant human interleukin 12 (rhIL-12).In some embodiments, an IL-15 based immunotherapeutic is heterodimericIL-15 (hetIL-15, Novartis/Admune), a fusion complex composed of asynthetic form of endogenous IL-15 complexed to the soluble IL-15binding protein IL-15 receptor alpha chain (IL15:sIL-15RA), which hasbeen tested in Phase 1 clinical trials for melanoma, renal cellcarcinoma, non-small cell lung cancer and head and neck squamous cellcarcinoma (NCT02452268). In some embodiments, a recombinant humaninterleukin 12 (rhIL-12) is NM-IL-12 (Neumedicines, Inc.), NCT02544724,or NCT02542124.

In some embodiments, an immuno-oncology agent is selected from thosedescripted in Jerry L. Adams ET. AL., “Big opportunities for smallmolecules in immuno-oncology,” Cancer Therapy 2015, Vol. 14, pages603-622, the content of which is incorporated herein by reference in itsentirety. In some embodiments, an immuno-oncology agent is selected fromthe examples described in Table 1 of Jerry L. Adams ET. AL. In someembodiments, an immuno-oncology agent is a small molecule targeting animmuno-oncology target selected from those listed in Table 2 of Jerry L.Adams ET. AL. In some embodiments, an immuno-oncology agent is a smallmolecule agent selected from those listed in Table 2 of Jerry L. AdamsET. AL.

In some embodiments, an immuno-oncology agent is selected from the smallmolecule immuno-oncology agents described in Peter L. Toogood, “Smallmolecule immuno-oncology therapeutic agents,” Bioorganic & MedicinalChemistry Letters 2018, Vol. 28, pages 319-329, the content of which isincorporated herein by reference in its entirety. In some embodiments,an immuno-oncology agent is an agent targeting the pathways as describedin Peter L. Toogood.

In some embodiments, an immuno-oncology agent is selected from thosedescribed in Sandra L. Ross et al., “Bispecific T cell engager (BiTE)antibody constructs can mediate bystander tumor cell killing”, PLoS ONE12(8): e0183390, the content of which is incorporated herein byreference in its entirety. In some embodiments, an immuno-oncology agentis a bispecific T cell engager (BiTE®) antibody construct. In someembodiments, a bispecific T cell engager (BiTE) antibody construct is aCD19/CD3 bispecific antibody construct. In some embodiments, abispecific T cell engager (BiTE®) antibody construct is an EGFR/CD3bispecific antibody construct. In some embodiments, a bispecific T cellengager (BiTE®) antibody construct activates T cells. In someembodiments, a bispecific T cell engager (BiTE) antibody constructactivates T cells, which release cytokines inducing upregulation ofintercellular adhesion molecule 1 (ICAM-1) and FAS on bystander cells.In some embodiments, a bispecific T cell engager (BiTE®) antibodyconstruct activates T cells which result in induced bystander celllysis. In some embodiments, the bystander cells are in solid tumors. Insome embodiments, the bystander cells being lysed are in proximity tothe BiTE®-activated T cells. In some embodiment, the bystander cellscomprises tumor-associated antigen (TAA) negative cancer cells. In someembodiment, the bystander cells comprise EGFR-negative cancer cells. Insome embodiments, an immuno-oncology agent is an antibody which blocksthe PD-L1/PD1 axis and/or CTLA4. In some embodiments, an immuno-oncologyagent is an ex-vivo expanded tumor-infiltrating T cell. In someembodiments, an immuno-oncology agent is a bispecific antibody constructor chimeric antigen receptors (CARs) that directly connect T cells withtumor-associated surface antigens (TAAs).

In certain embodiments, a combination of 2 or more therapeutic agentsmay be administered together with compounds of the invention. In certainembodiments, a combination of 3 or more therapeutic agents may beadministered with compounds of the invention.

Other examples of agents the inhibitors of this invention may also becombined with include, without limitation: vitamins and nutritionalsupplements, cancer vaccines, treatments for neutropenia (e.g. G-CSF,filgrastim, lenograstim), treatments for thrombocytopenia (e.g. bloodtransfusion, erythropoietin), PI3 kinase (PI3K) inhibitors, MEKinhibitors, mTOR inhibitors, CPT1 inhibitors, AMPK activators, PCSK9inhibitors, SREBP site 1 protease inhibitors, HMG CoA-reductaseinhibitors, antiemetics (e.g. 5-HT₃ receptor antagonists, dopamineantagonists, NK1 receptor antagonists, histamine receptor antagonists,cannabinoids, benzodiazepines, or anticholinergics), treatments forAlzheimer's Disease such as Aricept® and Excelon®; treatments forParkinson's Disease such as L-DOPA/carbidopa, entacapone, ropinrole,pramipexole, bromocriptine, pergolide, trihexephendyl, and amantadine;agents for treating Multiple Sclerosis (MS) such as beta interferon(e.g., Avonex® and Rebif®), Copaxone®, and mitoxantrone; treatments forasthma such as albuterol and Singulair®; agents for treatingschizophrenia such as zyprexa, risperdal, seroquel, and haloperidol;anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1 RA,azathioprine, cyclophosphamide, and sulfasalazine; immunomodulatory andimmunosuppressive agents such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, interferons, corticosteroids, cyclophophamide,azathioprine, and sulfasalazine; neurotrophic factors such asacetylcholinesterase inhibitors, MAO inhibitors, interferons,anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonianagents; agents for treating cardiovascular disease such asbeta-blockers, ACE inhibitors, diuretics, nitrates, calcium channelblockers, and statins, fibrates, cholesterol absorption inhibitors, bileacid sequestrants, and niacin; agents for treating liver disease such ascorticosteroids, cholestyramine, interferons, and anti-viral agents;agents for treating blood disorders such as corticosteroids,anti-leukemic agents, and growth factors; agents for treatingimmunodeficiency disorders such as gamma globulin; and anti-diabeticagents such as biguanides (metformin, phenformin, buformin),thiazolidinediones (rosiglitazone, pioglitazone, troglitazone),sulfonylureas (tolbutamide, acetohexamide, tolazamide, chlorpropamide,glipizide, glyburide, glimepiride, gliclazide), meglitinides(repaglinide, nateglinide), alpha-glucosidase inhibitors (miglitol,acarbose), incretin mimetics (exenatide, liraglutide, taspoglutide),gastric inhibitory peptide analogs, DPP-4 inhibitors (vildagliptin,sitagliptin, saxagliptin, linagliptin, alogliptin), amylin analogs(pramlintide), and insulin and insulin analogs.

In certain embodiments, compounds of the present invention, or apharmaceutically acceptable composition thereof, are administered incombination with antisense agents, a monoclonal or polyclonal antibodyor an siRNA therapeutic.

In another embodiment, the present invention provides a method oftreating an inflammatory disease, disorder or condition by administeringto a patient in need thereof a compound of the present invention and oneor more additional therapeutic agents. Such additional therapeuticagents may be small molecules or recombinant biologic agents andinclude, for example, acetaminophen, non-steroidal anti-inflammatorydrugs (NSAIDS) such as aspirin, ibuprofen, naproxen, etodolac (Lodine®)and celecoxib, colchicine (Colcrys®), corticosteroids such asprednisone, prednisolone, methylprednisolone, hydrocortisone, and thelike, probenecid, allopurinol, febuxostat (Uloric®), sulfasalazine(Azulfidine®), antimalarials such as hydroxychloroquine (Plaquenil®) andchloroquine (Aralen®), methotrexate (Rheumatrex®), gold salts such asgold thioglucose (Solganal®), gold thiomalate (Myochrysine®) andauranofin (Ridaura®), D-penicillamine (Depen® or Cuprimine®),azathioprine (Imuran®), cyclophosphamide (Cytoxan®), chlorambucil(Leukeran®), cyclosporine (Sandimmune®), leflunomide (Arava®) and“anti-TNF” agents such as etanercept (Enbrel®), infliximab (Remicade®),golimumab (Simponi®), certolizumab pegol (Cimzia®) and adalimumab(Humira®), “anti-IL-1” agents such as anakinra (Kineret®) and rilonacept(Arcalyst®), canakinumab (Ilaris®), anti-Jak inhibitors such astofacitinib, antibodies such as rituximab (Rituxan®), “anti-T-cell”agents such as abatacept (Orencia®), “anti-IL-6” agents such astocilizumab (Actemra®), diclofenac, cortisone, hyaluronic acid (Synvisc®or Hyalgan®), monoclonal antibodies such as tanezumab, anticoagulantssuch as heparin (Calcinparine® or Liquaemin®) and warfarin (Coumadin®),antidiarrheals such as diphenoxylate (Lomotil®) and loperamide(Imodium®), bile acid binding agents such as cholestyramine, alosetron(Lotronex®), lubiprostone (Amitiza®), laxatives such as Milk ofMagnesia, polyethylene glycol (MiraLax®), Dulcolax®, Correctol® andSenokot®, anticholinergics or antispasmodics such as dicyclomine(Bentyl®), Singulair®, beta-2 agonists such as albuterol (Ventolin® HFA,Proventil® HFA), levalbuterol (Xopenex®), metaproterenol (Alupent®),pirbuterol acetate (Maxair®), terbutaline sulfate (Brethaire®),salmeterol xinafoate (Serevent®) and formoterol (Foradil®),anticholinergic agents such as ipratropium bromide (Atrovent®) andtiotropium (Spiriva®), inhaled corticosteroids such as beclomethasonedipropionate (Beclovent®, Qvar®, and Vanceril®), triamcinolone acetonide(Azmacort®), mometasone (Asthmanex®), budesonide (Pulmocort®), andflunisolide (Aerobid®), Afviar®, Symbicort®, Dulera®, cromolyn sodium(Intal®), methylxanthines such as theophylline (Theo-Dur®, Theolair®,Slo-bid®, Uniphyl®, Theo-24®) and aminophylline, IgE antibodies such asomalizumab (Xolair®), nucleoside reverse transcriptase inhibitors suchas zidovudine (Retrovir®), abacavir (Ziagen®), abacavir/lamivudine(Epzicom®), abacavir/lamivudine/zidovudine (Trizivir®), didanosine(Videx®), emtricitabine (Emtriva®), lamivudine (Epivir®),lamivudine/zidovudine (Combivir®), stavudine (Zerit®), and zalcitabine(Hivid®), non-nucleoside reverse transcriptase inhibitors such asdelavirdine (Rescriptor®), efavirenz (Sustiva®), nevairapine (Viramune®)and etravirine (Intelence®), nucleotide reverse transcriptase inhibitorssuch as tenofovir (Viread®), protease inhibitors such as amprenavir(Agenerase®), atazanavir (Reyataz®), darunavir (Prezista®),fosamprenavir (Lexiva®), indinavir (Crixivan®), lopinavir and ritonavir(Kaletra®), nelfinavir (Viracept®), ritonavir (Norvir®), saquinavir(Fortovase® or Invirase®), and tipranavir (Aptivus®), entry inhibitorssuch as enfuvirtide (Fuzeon®) and maraviroc (Selzentry®), integraseinhibitors such as raltegravir (Isentress®), doxorubicin(Hydrodaunorubicin®), vincristine (Oncovin®), bortezomib (Velcade®), anddexamethasone (Decadron®) in combination with lenalidomide (Revlimid®),or any combination(s) thereof.

In some embodiments, a provided compound is administered in combinationwith an antiviral agent, including, e.g., acyclovir, pencyclovir,cidofovir, idoxuridine, zidovudine, ribavarin, amantadine, foscarnet,didanosine, acyclovir, ganciclovir, cidofovir, zalcitabine, rimantadine,calacyclovir, famiciclovir, abacavir, didanosine, emtricitabine,lamivudine, stavudine, tenofovir, zalcitabine, zidovudine,zidovudine-lamivudine, TRIZIVIR (zidovudine, lamivudine, abacavir),EPZICOM (aba-cavir-lamivudine), TRUVADA (tenofovir-emtricitabine),efavirenz, nevirapine, and delavirdine, amprenavir, atazanavir,fosamprenavir, indinavir, lopinavir-ritonavir, nelfinavir, ritonavir,saquinavir, and tipranavir. In some embodiments, the antiviral agent isanti-influenza agent including, e.g., rimantadine, amantadine,oseltamivir, and zanamivir.

Those additional agents may be administered separately from an inventivecompound-containing composition, as part of a multiple dosage regimen.Alternatively, those agents may be part of a single dosage form, mixedtogether with a compound of this invention in a single composition. Ifadministered as part of a multiple dosage regime, the two active agentsmay be submitted simultaneously, sequentially or within a period of timefrom one another, normally within five hours from one another.

As used herein, the term “combination,” “combined,” and related termsrefers to the simultaneous or sequential administration of therapeuticagents in accordance with this invention. For example, a compound of thepresent invention may be administered with another therapeutic agentsimultaneously or sequentially in separate unit dosage forms or togetherin a single unit dosage form. Accordingly, the present inventionprovides a single unit dosage form comprising a compound of the presentinvention, an additional therapeutic agent, and a pharmaceuticallyacceptable carrier, adjuvant, or vehicle.

The amount of both, a provided compound and additional therapeutic agent(in those compositions which comprise an additional therapeutic agent asdescribed above) that may be combined with the carrier materials toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. Preferably, compositions ofthis invention should be formulated so that a dosage of between 0.01-100mg/kg body weight/day of an inventive can be administered.

In those compositions which comprise an additional therapeutic agent,that additional therapeutic agent and the compound of this invention mayact synergistically. Therefore, the amount of additional therapeuticagent in such compositions will be less than that required in amonotherapy utilizing only that therapeutic agent. In such compositionsa dosage of between 0.01-100 μg/kg body weight/day of the additionaltherapeutic agent can be administered.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered in a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

In one embodiment, the present invention provides a compositioncomprising a compound of the present invention and one or moreadditional therapeutic agents. The therapeutic agent may be administeredtogether with a compound of the present invention, or may beadministered prior to or following administration of a compound of thepresent invention. Suitable therapeutic agents are described in furtherdetail below. In certain embodiments, a compound of the presentinvention may be administered up to 5 minutes, 10 minutes, 15 minutes,30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5, hours, 6 hours, 7hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14hours, 15 hours, 16 hours, 17 hours, or 18 hours before the therapeuticagent. In other embodiments, a compound of formula I′ may beadministered up to 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1hour, 2 hours, 3 hours, 4 hours, 5, hours, 6 hours, 7 hours, 8 hours, 9hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16hours, 17 hours, or 18 hours following the therapeutic agent.

In some embodiments, the present invention provides a medicamentcomprising at least one compound of the present invention or apharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable carrier.

EXEMPLIFICATION

As depicted in the Examples below, in certain exemplary embodiments,compounds are prepared according to the following general procedures. Itwill be appreciated that, although the general methods depict thesynthesis of certain compounds of the present invention, the followinggeneral methods, and other methods known to one of ordinary skill in theart, can be applied to all compounds and subclasses and species of eachof these compounds, as described herein.

Example 1A

DRE-Luciferase Reporter Assay

AHR binds to Dioxin Responsive Elements (DRE) upstream of genes that itactivates. One measure of AHR activity is activation of a reporter gene,such as luciferase, downstream of one or multiple DRE elements.Luciferase activity will reflect activation and inhibition of AHR in thecells expressing his reporter.

Murine Hepa1-6 or Hepa-1c1c7 or other murine cell line with aDRE-luciferase reporter either stably or transiently transfected wereplated in media in plates (96-well, 384-well or other plates) andincubated overnight at 37° C. in a CO₂ incubator. Likewise, human HepG2or other human cell line with a DRE-luciferase reporter either stably ortransiently transfected were plated in media in plates (96-well,384-well or other plates) and incubated overnight at 37° C. in a CO₂incubator.

The next day, an AHR activating ligand, such as TCDD, kynurenine, ITE(2-(1H-indole-3-ylcarbonyl)-4-thiazolecarboxylic methyl ester), VAF347,BNF (beta-naphthoflavone), FICZ (6-formylindolo(3,2-b) carbazole orother AHR ligands, was added with or without AHR antagonist.

Cells were incubated for 4, 15 or 24 hours or another time point andthen lysed for determination of luciferase activity as a read-out of theAHR activation or inhibition. Luciferase was measured with a commercialkit such as the Promega Luciferase kit or any kit or reagents thatprovide the luciferin substrate for measuring luciferase activity. Thelevel of luciferase with only activating ligand added was the maximumsignal while the luciferase with no ligand was the minimum signal. IC₅₀values were determined as the concentration which inhibits half of theluciferase activity. Compounds assayed and their IC₅₀ values are shownin Table 2, below.

In some embodiments, compounds have an IC₅₀ of 5-20 μM. In someembodiments, compounds have an IC₅₀≤5 μM. In some embodiments, compoundshave an IC₅₀≤1 μM. In some embodiments, compounds have an IC₅₀≤0.1 μM.In some embodiments, compounds have an IC₅₀≤0.01 μM. In someembodiments, compounds have an IC₅₀≤0.001 μM.

Activity of certain compounds of the present invention as obtained bythe above assay is set forth in Table 2, below.

In Table 2, IC50 values are reported as A, B, C and D, whereby Arepresents an IC50 of <0.5 μM; B represents an IC50 of between 0.5 and1.0 μM; and C represents an IC50 of between 1.0 and 1.5 μM; and Drepresents an IC50 of >1.5 μM.

TABLE 2 IC₅₀ Values for Select Compounds Assayed According to Example1A. Compound IC₅₀ I-1 C I-2 A I-3 C I-4 A I-5 D I-6 A I-7 A I-9 C  I-19B

Example 1B

DRE-Luciferase Reporter Assay (Alternate Method)

AHR binds to Dioxin Responsive Elements (DRE) upstream of genes that itactivates. One measure of AHR activity is activation of a reporter gene,such as luciferase, downstream of one or multiple DRE elements.Luciferase activity will reflect activation and inhibition of AHR in thecells expressing his reporter.

Murine Hepa1-6 or Hepa-1c1c7 or other murine cell line with aDRE-luciferase reporter either stably or transiently transfected wereplated in media in plates (96-well, 384-well or other plates) andincubated overnight at 37° C. in a CO₂ incubator or compound and agonistwere added at the time of plating. Likewise, human HepG2 or other humancell line with a DRE-luciferase reporter either stably or transientlytransfected were plated in media in plates (96-well, 384-well or otherplates) and incubated overnight at 37° C. in a CO₂ incubator or compoundand agonist were added at the time of plating.

At the time that cells are plated or following incubation overnight, anAHR activating ligand, such as TCDD, kynurenine, ITE(2-(1H-indole-3-ylcarbonyl)-4-thiazolecarboxylic methyl ester), VAF347,BNF (beta-naphthoflavone), FICZ (6-formylindolo(3,2-b) carbazole orother AHR ligands, was added with or without AHR antagonist.

Cells were incubated for 4, 15 or 24 hours or another time point andthen lysed for determination of luciferase activity as a read-out of theAHR activation or inhibition. Luciferase was measured with a commercialkit such as the Promega Luciferase kit or any kit or reagents thatprovide the luciferin substrate for measuring luciferase activity. Thelevel of luciferase with only activating ligand added was the maximumsignal while the luciferase with no ligand was the minimum signal. IC₅₀values were determined as the concentration which inhibits half of theluciferase activity. Compounds assayed and their IC₅₀ values are shownin Table 3, below.

In some embodiments, compounds have an IC₅₀ of 5-20 μM. In someembodiments, compounds have an IC₅₀≤5 μM. In some embodiments, compoundshave an IC₅₀≤1 μM. In some embodiments, compounds have an IC₅₀≤0.1 μM.In some embodiments, compounds have an IC₅₀≤0.01 μM. In someembodiments, compounds have an IC₅₀≤0.001 μM.

Activity of certain compounds of the present invention as obtained bythe above assay is set forth in Table 3, below.

In Table 3, IC₅₀ values are reported as A, B, C and D, whereby Arepresents an IC₅₀ of <0.5 μM; B represents an IC₅₀ of between 0.5 and1.0 μM; and C represents an IC₅₀ of between 1.0 and 1.5 μM; and Drepresents an IC₅₀ of >1.5 μM.

TABLE 3 IC₅₀ Values for Select Compounds Assayed According to Example1B. Compound IC₅₀ Compound IC₅₀ I-1 A I-58 D I-2 A I-59 A I-3 A I-60 BI-4 A I-61 A I-5 A I-62 A I-6 A I-63 A I-7 D I-64 A I-8 D I-65 A I-9 BI-66 A I-10 D I-67 A I-11 A I-68 B I-13 D I-69 A I-14 D I-70 A I-15 AI-71 A I-16 A I-72 A I-19 A I-73 A I-20 A I-74 B I-21 A I-75a D I-22 AI-75b D I-23 A I-77 A I-24 D I-78 A I-25 D I-79 A I-26 D I-80 B I-27a BI-81 A I-28a D I-82b D I-29 D I-83 A I-30 A I-84 D I-31 D I-85 B I-32a DI-86 A I-33 A I-87 A I-34 D I-88 B I-35 D I-89 A I-36 D I-90 A I-37a DI-91 A I-38 D I-92 A I-39 A I-93 A I-40 A I-94a D I-41 D I-95 D I-42a DI-96 A I-43 B I-97a D I-44 D I-98a D I-45 A I-99 A I-46 A I-100 D I-47 AI-101 A I-48 A I-102 A I-49 A I-103 A I-50 D I-104 B I-51 A I-105 A I-52B I-106 A I-53 A I-107 A I-54 A I-108 A I-55 A I-109 A I-56 A I-110 BI-57 A I-111 A I-120 D I-185 D I-121 D I-186 B I-122 D I-187a D I-123 CI-187b D I-124 D I-125 D I-189 D I-126 A I-190a D I-128a D I-190b DI-128b D I-192 B I-129 A I-193 D I-130 A I-194 D I-131 A I-195 D I-132 BI-196 A I-133 A I-197 A I-134 D I-198a A I-135 A I-198b B I-136 A I-200A I-137a A I-201 A I-137b A I-202 A I-139a A I-203 A I-139b A I-204a AI-141 A I-204b A I-142 D I-206 D I-143 D I-207 D I-144a C I-208 D I-144bA I-209 B I-146a D I-210 C I-146b D I-211 D I-148 D I-212 D I-149 DI-213 A I-150a A I-214 A I-150b C I-215 D I-152a D I-216a B I-152b AI-216b D I-154 A I-218 A I-155 B I-219 A I-156 D I-220 A I-157 D I-221 DI-158a D I-222a A I-158b D I-222b A I-160a D I-224 A I-160b D I-225 AI-162 A I-226 A I-163 A I-227 A I-164 A I-228 D I-165 A I-229 D I-166 AI-230 A I-167 D I-231 D I-168 D I-232 A I-169 B I-233 B I-170 D I-234 BI-171 C I-235 A I-172 C I-236 D I-173 D I-237 A I-174 D I-238 D I-175 DI-239 D I-176 D I-240 D I-177 A I-241 A I-178 D I-242 A I-179 A I-243a DI-180 B I-243b D I-181 D I-245 D I-182 D I-246 C I-183 D I-247 D I-184 DI-248 D I-249a A I-249b A I-250 D I-251 D I-252a A I-252b A I-253 AI-254 D I-255 A I-256 A I-257 A I-258 D I-259a A I-259b A I-260 D I-261D I-262 A I-263a C I-263b C I-264 D I-265 A I-266 D I-267 A I-268 DI-269 D I-270 A I-271 D I-272 D I-273 C I-274 A I-275 A I-276 A I-277 AI-278 A I-279 D I-280 D I-281 A I-282 A I-283 D I-284 B I-285 D I-286 AI-287 D I-288 A I-289 A

Example 1C

Mouse Pharmacokinetic Study

Formulations of compounds set forth in Table 4 were administeredintravenously or orally via gavage to CD-1 mice. Typically, at 0.167,0.5, 1, 2, 4, 6, 12, and 24 hours post-dose, blood was collected andprocessed to plasma by centrifugation and stored at −80° C. untilanalysis.

Internal standard was added to each sample prior to proteinprecipitation with acetonitrile. The precipitates were filtered througha Phree phospholipid removal filter plate and the samples were analyzedby LC/MS/MS. A standard curve was prepared in plasma from typically from1.0 ng/mL to 3000 ng/mL and processed in the same manner as the samples.Sample analysis was typically performed on a suitable LC/MS/MS systemfitted with an analytical UPLC column and compounds eluted from theanalytical column with a gradient from 30-95% 0.1% formic acid (v/v) inACN: 0.1% formic acid (v/v) in water. Mass spectrometric detection oftest compound and the internal standard was performed by MRM in positivemode. The pharmacokinetics of each compound were analyzed by PhoenixWinNonlin software (Pharsight, St. Louis, Mo.) via non-compartmentalanalysis. The results are summarized in Table 4 below.

TABLE 4 Mouse Pharmacokinetic Data According to Example 1C. Cl AUC AUCBio- Dose C_(max) Vdss (mL/ 0-last 0-inf avail- (mg/ (ng/ T_(max)T^(1/2) (L/ min/ T_(last) (ng * h/ (ng * h/ ability Compound kg) RouteFormulation mL) (h) (h) kg) kg) (h) mL) mL) (%) I-137a 3 iv 1.5 mg/mL8.2 6.6 25 24 1921 2008 in DMSO:PEG 400:Water = 5:75:20, solution I-137a30 po 3 mg/mL in 953 0.5 3.5 24 3782 3835 19 0.5% methyl- cellulose and0.2% Tween 80, suspension I-137b 3 iv 1.5 mg/mL 11.2 16.6 37 24 12201352 in DMSO:PEG 400:Water = 5:75:20, solution I-137b 30 po 3 mg/mL in476 2 5.9 24 3630 3828 28 0.5% methyl- cellulose and 0.2% Tween 80,suspension I-144b 30 po 1.5 mg/mL 1.3 0.8 44 6 1140 1145 in DMSO:PEG400:Water = 5:75:20, solution I-144b 30 po 3 mg/mL in 599 0.2 2 6 613727 6 0.5% methyl- cellulose and 0.2% Tween 80, suspension I-154 3 iv1.5 mg/mL 12 13.7 16.5 24 2414 3034 in DMSO:PEG 400:Water = 5:75:20,solution I-154 30 po 3 mg/mL in 840 2 9.6 24 9358 11207 39 0.5% methyl-cellulose and 0.2% Tween 80, suspension I-198a 3 iv 1.5 mg/mL 4.3 12.164.8 24 765 772 in DMSO:PEG 400:Water = 5:75:20, solution I-198a 30 po3.00 mg/mL 721 1 4.2 24 3961 4048 52 in 0.5% Methyl- cellulose,suspension I-201 3 iv 1.5 mg/mL 8.52 19.6 37.5 24 1203 1334 in DMSO:PEG400:Water = 5:75:20, solution I-201 30 po 3.00 mg/mL 1413 1 7.16 24 965310558 79 in 0.5% Methyl- cellulose, suspension I-203 3 iv 1.5 mg/mL 2.35.8 88.5 12 555 565 in DMSO:PEG 400:Water = 5:75:20, solution I-203 30po 3.00 mg/mL 604 0.2 0.7 6 656 657 12 in 0.5% Methyl- cellulose,suspension I-204b 3 iv 1.5 mg/mL 0.9 4 110 6 454 455 in DMSO:PEG400:Water = 5:75:20, solution I-204b 30 po 3.00 mg/mL 55 0.2 3.2 12 6367 2 in 0.5% Methyl- cellulose, suspension I-219 3 iv 1.5 mg/mL 0.7 2.473.3 6 678 682 in DMSO:PEG 400:Water = 5:75:20, solution I-219 30 po3.00 mg/mL 48.9 0.2 12 132 215 2 in 30% PEG400/ 10% solutol/ 60% water,suspension I-222a 3 iv 1.5 mg/mL 2.4 1.5 36.9 6 1329 1329 in DMSO:PEG400:Water = 5:75:20, solution I-222a 30 po 3.00 mg/mL 2873 0.2 1.7 122231 2239 16 in 30% PEG400/ 10% solutol/ 60% water, solution I-222b 3 iv1.5 mg/mL 1.8 2 88.1 6 564 567 in DMSO:PEG 400:Water = 5:75:20, solutionI-222b 30 po 3.00 mg/mL 2167 0.2 3.3 24 2865 2870 51 in 30% PEG400/ 10%solutol/ 60% water, solution I-39 3 iv 1.5 mg/mL 1.31 3.5 53.7 6 908 931in DMSO:PEG 400:Water = 5:75:20, solution I-39 30 po 3.00 mg/mL 799 3.3524 1597 1609 17 in DMSO:PEG 400:Water = 5:75:20, solution I-40 3 iv 1.5mg/mL 8.4 24 41.2 24 1066 1213 in DMSO:PEG 400:Water = 5:75:20, solutionI-40 30 po 3.00 mg/mL 853 2 8.8 24 7560 9142 75 in 30% PEG400/ 10%solutol/ 60% water, solution I-65 3 iv 1.5 mg/mL 0.7 2.7 93.1 4 522 537in DMSO:PEG 400:Water = 5:75:20, solution I-65 30 po 3.00 mg/mL 574 0.20.8 6 370 373 7 in DMSO:PEG 400:Water = 5:75:20, solution I-70 3 iv 1.5mg/mL 1.08 3.2 54.9 6 862 872 in DMSO:PEG 400:Water = 5:75:20, solutionI-70 30 po 10.0 mg/mL 900 0.5 2.26 12 1944 1993 26 in 30% PEG/ 70% (20%hp-?-CD) solution, clear solution I-73 3 iv 1.5 mg/mL 2 7.7 90 12 553556 in DMSO:PEG 400:Water = 5:75:20, solution I-73 30 po 3.00 mg/mL 37.51 2.1 6 24 96 108 2 in 0.5% Methyl- cellulose, suspension I-75a 3 iv 1.5mg/mL 13.8 13.7 14.4 24 2666 2265 in DMSO:PEG 400:Water = 5:75:20,solution I-75a 30 po 3.0 mg/mL 969 2 9.8 24 13579 936 51 in DMSO:PEG400:Water = 5:75:20, solution I-103 3 iv 3.00 mg/mL 0.62 1.4 43.9 6 11371140 in 40% PEG400/ 10% solutol/ 50% water, solution I-103 30 po 6.00mg/mL 713 0.2 2.15 12 649 671 6 in 40% PEG400/ 10% solutol/ 50% water,solution I-107 3 iv 3.00 mg/mL 2.18 43.2 540 6 88 92 in DMSO:solutol:water = 5:10:85, solution I-107 15 po 3.00 mg/mL 0 0 0 0 0 0 inDMSO: solutol:water = 5:10:85, solution I-2 3 iv 3.00 mg/mL 1.99 9.6 826 554 610 in DMSO:PEG 400:Water = 5:75:20, solution I-2 30 po 6.00 mg/mL182 0.2 3.9 24 730 743 12 in DMSO:PEG 400:Water = 5:75:20, solutionI-259a 3 iv 1.5 mg/mL 1.2 4.7 59 6 825 842 in DMSO:PEG 400:Water =5:75:20, clear solution I-259a 30 po 3.0 mg/mL 82 0.5 1.2 6 237 249 3 in0.5% methyl- cellulose, homogenous opaque suspension I-118 3 iv 1.5mg/mL 0.6 4.9 136 4 364 368 in DMSO:PEG 400:Water = 5:75:20, clearsolution I-118 30 po 3.0 mg/mL 41 1 1.9 6 107 121 3 in 0.5% methyl-cellulose, homogenous opaque suspension I-126 3 iv 1.5 mg/mL 8.8 17 3024 1463 1651 in DMSO:PEG 400:Water = 5:75:20, clear solution I-126 30 po3.0 mg/mL 1870 1 6.1 24 13748 14741 100 in 0.5% methyl- cellulose,homogenous opaque suspension I-129 3 iv 1.5 mg/mL 14 22 31 24 1339 1615in DMSO:PEG 400:Water = 5:75:20, clear solution I-129 30 po 3.0 mg/mL1043 2 6.4 24 7034 7532 53 in 0.5% methyl- cellulose, homogenous opaquesuspension I-130 3 iv 1.5 mg/mL 2.6 7.5 39 12 1223 1272 in DMSO:PEG400:Water = 5:75:20, clear solution I-130 30 po 3.0 mg/mL 936 2 3.5 246624 6689 53 in 0.5% methyl- cellulose, homogenous opaque suspensionI-59 3 iv 1.5 mg/mL 6.6 11.2 53 24 925 949 in DMSO:PEG 400:Water =5:75:20, clear solution I-59 30 po 3.0 mg/mL 719 0.5 4.3 24 2110 2130 22in 0.5% methyl- cellulose, homogenous opaque suspension

Example 1D

In Vitro Mouse liver S9 Metabolic Stability Assay

CD-1 mouse liver S9 were purchased from Corning or XenoTech LLC orBioreclamationIVT, LLC or WuXi prepared. The cells were stored at −80°C. in a freezer before use. β-Nicotinamide adenine dinucleotidephosphate (NADP), Glucose 6-phosphate (G6P), Glucose 6-phosphatedehydrogenase from yeast (G6PDH), Uridine 5′-diphophoglucuronic acidtrisodium salt (UDPGA) and Adenosine 3′-phosphate 5′-phosphosulfatelithium salt hydrate (PAPS) were available commercially from Sigma.

Compounds were diluted in DMSO to make 10 mM stock solution. 5 μL ofthis stock solution (10 mM, DMSO) was diluted with 45 μL DMSO and 450 μL50% Methanol/Water to make intermediate stock solution (100 μM, 45%MeOH, 10% DMSO). 50 μL of intermediate stock solution was diluted with450 μL 100 mM phosphate buffer to make a final stock solution (10 μM,4.5% MeOH, 1% DMSO). 10 uL of final stock solution was added to 90 uLliver S9 system (final concentration of 1 μM, 0.45% MeOH, 0.1% DMSO).

Test compounds were incubated at 37° C. with liver S9 (pooled frommultiple donors) at 1 μM in the presence of a NADPH regenerating system,UDPGA, and PAPS at 1 mg/mL S9 protein. Time samples (0 and 60 minutes)were removed and immediately mixed with cold acetonitrile containinginternal standard (IS). Samples were analyzed by LC/MS/MS anddisappearance of test compounds were assessed based on peak area ratiosof analyte /IS (no standard curve). All samples were injected andanalyzed using LC-MS/MS. The analyte/internal standard peak area ratioswere converted to percentage remaining (% Remaining) with the followingequation: % Remaining at 60 min=(Peak area ratio of analyte to IS at 60min/Peak area ratio of analyte to IS at t=0)×100%. The results aresummarized in Table 5 below. Table 5. Mouse Stability Data According toExample 1D.

TABLE 5 Mouse Stability Data According to Example 1D. Mouse S9Stability: Mouse S9 Stability: % Remaining After % Remaining AfterCompound 60 Minutes Compound 60 Minutes I-6 0 I-40 87 I-2 13 I-39 30 I-40 I-36 29 I-21 26 I-30 11 I-16 2 I-235 2 I-109 10 I-224 0 I-107 0 I-222b0 I-106 1 I-222a 1 I-105 8 I-219 5 I-103 7 I-214 0 I-101 1 I-209 0 I-98b0 I-208 0 I-96 36 I-204b 0 I-92 0 I-203 0 I-90 11 I-202 0 I-81 2 I-20134 I-78 1 I-200 0 I-75a 65 I-198a 30 I-73 11 I-196 0 I-72 3 I-177 2 I-700 I-166 4 I-68 4 I-165 3 I-67 5 I-164 0 I-67 1 I-162 54 I-65 0 I-156 28I-59 11 I-154 13 I-47 50 I-152b 3 I-45 19 I-152a 2

Example 1E

In Vivo Mouse liver and spleen Cyp1a1 Modulation Assay

C57BL/6 mice, female, 6-8 weeks old, weighing approximately 18-20 g werepurchased from Shanghai Lingchang Biological Technology Co., Ltd orother certified vendors and used in the studies. Animal husbandry,feeding and health conditions are according to animal welfareguidelines. VAG539 (30 mg/kg, po) was used as AHRagonist, and testcompounds were formulated in suitable vehicles, typically 0.5%methylcellulose).

C57BL/6 mice (n=3 per group) were treated with AHR agonist alone or withAHR agonist and test compounds. Animals were sacrificed at 4 or 10 hoursafter treatment upon which their livers and spleens were collected andsubsequently analyzed by qPCR. Normalized fold induction of cyp1a1 wasdetermined by comparing mCYP1A1 and mGAPDH counts (ct) according to:normalized fold=2^(−ΔΔCt). The percent inhibition was calculatedaccording to:

${\left\lbrack {1 - \frac{\begin{matrix}{{{average}\mspace{14mu}{norma1ized}\mspace{14mu}{fold}\mspace{14mu}{for}}\mspace{14mu}} \\{{AHR}\mspace{14mu}{agonist}\mspace{14mu}{and}\mspace{14mu}{compound}\mspace{14mu}{treated}}\end{matrix}}{{average}\mspace{14mu}{norma1ized}\mspace{14mu}{fold}\mspace{14mu}{for}\mspace{14mu}{AHR}\mspace{14mu}{agonist}\mspace{14mu}{treated}}} \right\rbrack \times 100} = {\%\mspace{14mu}{inhibition}}$The results are summarized in Table 6 below.

TABLE 6 Cyp1a1 Inhibition Data According to Example 1E. Cyp1a1inhibition Liver Spleen Compound Dose 4 h 10 h 4 h I-2 30 mg/kg 50%I-103 30 mg/kg 94% 0% I-75a 30 mg/kg 55% I-57 30 mg/kg 77% 66% I-46 30mg/kg 96% 94% I-39 30 mg/kg 90% 69% I-70 30 mg/kg 98% 10 mg/kg 99% 81%10 mg/kg 98% 98% 88% 5 mg/kg 93% 96% 79% 2 mg/kg 83% 63% 65% 1 mg/kg 52%34% 38% I-40 25 mg/kg 98% 97% 96% 10 mg/kg 93% 75% 74% 5 mg/kg 77% 48%34% I-201 25 mg/kg 86% 97% 72% 10 mg/kg 72% 59% 46% 5 mg/kg 58% 44% 40%

Example 1F

T-Cell Study with I-70

Human T cells were isolated by CD3 negative selection after isolation ofPBMCs from blood of human donors via ficoll density gradientcentrifugation. One million T cells were activated with 25 uL ofCD3/CD28 tetramer (Stemcell) in the presence or absence of I-70 for 24hours, after which media was removed and stored at −80 C for latercytokine analysis. Cells were then washed 2× with PBS, before isolatingRNA according to the manufacturer's instructions for the RNAeasy minikit (Qiagen).

RNA was converted to cDNA using VILO-IV RT mastermix (Thermofisher), andq-RT-PCR was performed to determine levels of IL-22 (Hs01574154_m1),Cyp1a1 (Hs01054797_g1), and GAPDH (Hs00266705_g1). Data was analyzedusing the ddCT method whereby each sample is first normalized to GAPDHhousekeeping gene before being normalized to control treatment. IL22 andcyp1a1 RNA expression levels are inhibited by treatment with I-70, asshown in FIG. 1.

Cytokine levels were determined utilizing the mesoscale discovery (MSD)platform (K15067L-2) and MSD analysis software according to themanufacturer's instructions. IL-22 protein levels are decreased andpro-inflammatory IL-2 protein levels are increased by treatment withI-70, as shown in FIG. 2.

CD3/CD28 activated T cells are AHR activated as measured by geneexpression and cytokine production. Treatment with the AHR inhibitorlead to inhibition of cyp1a1 and IL22 gene expression and cytokine IL-22production. AHR inhibition also increases production of thepro-inflammatory cytokine IL-2.

Example 1G

Efficacy Study of I-70 and Checkpoint Inhibitor Anti-PD-1 in the MouseColorectal Cancer Model CT26 in Balb/c Mice

CT26 is a murine colon carcinoma cell line obtained from ATCC. CT26cells were cultured in RPMI supplemented with 10% FBS. 5×10⁵ CT26 cellsin 100 μl PBS were implanted subcutaneously in 6-8 week old female,Balb/c mice. Dosing for the efficacy study starts 4 days post implant:AHR antagonist was dosed orally, every day (QD) at or 10 mg/kg for 3weeks. anti-PD-1 (BioXcell RMP1-14) was twice a week, intraperitoneally(IP) at 10 mg/kg for five total doses. Tumors were monitored by calipermeasurement every 2-3 days and body weight measured three times perweek.

Tumor growth is inhibited by AHR antagonist I-70 alone or in combinationwith anti-PD-1, as shown in FIG. 3. Tumor growth inhibition isstatistically significant with I-70 as single agent compared to vehicle,with p value=0.0166. In addition, the tumor growth inhibition in thecombination group was significant compared to the anti-PD-1 alone, pvalue=0.0420. p values determined by Student's T-test analysis.

Example 1H

Efficacy Study of I-70 and Checkpoint Inhibitor Anti-PD-1 in the MouseMelanoma Model B16-IDO in C57BL/6 Mice

B16-IDO is a murine melanoma carcinoma cell line that has beenengineered to overexpress IDO1 (Holmgaard, 2015 Cell Reports). B16-IDOcells were cultured in DMEM supplemented with 10% FBS. 2×10⁵ B16-IDOcells in 50 μl PBS were implanted intradermally in 6-8 week old female,C57BL/6 mice. Dosing for the efficacy study starts 7 days post implant:AHR antagonist I-70 was dosed orally, every day (QD) at or 10 mg/kg for2 weeks. Anti-PD-1 (BioXcell RMP1-14) was administered every 3^(rd) day,intraperitoneally (IP) at 250 μg/mouse for five total doses. Tumors weremonitored by caliper measurement every 2-3 days and body weight measuredthree times per week.

Tumor growth was inhibited by AHR antagonist I-70 alone or incombination with anti-PD-1, as shown in FIG. 4. Tumor growth inhibitionwas statistically significant with I-70 as single agent compared tovehicle, with p value <0.001. In addition, the tumor growth inhibitionin the combination group was significant compared to the anti-PD-1alone, p value <0.03. p values determined by Student's T-test analysis.

Example 2

AHR-Dependent Gene Expression

Murine Hepa1-6 or Hepa-1c1c7 or other murine cell line are plated inmedia in plates (6, well, 12 well or other plates) and incubatedovernight at 37° C. in a CO₂ incubator; or human HepG2 or other humancell line are plated in media in plates (6-well, 12-well or otherplates) and incubated overnight at 37° C. in a CO₂ incubator.

The next day AHR activating ligand, such as TCDD, kynurenine, ITE(2-(1H-indole-3-ylcarbonyl)-4-thiazolecarboxylic methyl ester), VAF347,BNF (beta-naphthoflavone), ICZ (6-Formylindolo(3,2-b) carbazole or otherAHR ligands added with or without AHR antagonist. Cells are incubatedfor 4, 15 or 24 hours or another time point and then cells are lysed forRNA collection. RNA can be collected via a RNA isolation kit such asQiagen or any other RNA isolation method. Gene expression is determinedby quantitative RT-PCR using probes for specific genes including ahousekeeping gene such as Gapdh, β-actin or other constitutivelyexpressed genes for normalization. AHR-dependent genes to be examinedinclude but are not limited to: cyp1a1, cyp1b1, AHRR, IDO1, IDO2, cox2,IL6, VEGFA, cyclinD1, cdc2, MMP-9, c-myc.

Example 3

AHR-dependent gene expression is measured in tissue samples such astumor or liver. RNA is extracted from the tissue via methods such as RNAisolation kit such as Qiagen or any other RNA isolation method known toone of ordinary skill in the art. The RNA extraction could be done fromtotal cells or cells post-sorting for specific populations of cells suchas tumor cells, tumor associated-T cells, tumor associated-myeloid cellsor others. Gene expression is determined by quantitative RT-PCR usingprobes for specific genes including a housekeeping gene such as Gapdh,β-actin or other constitutively expressed genes for normalization.AHR-dependent genes to be examined include but are not limited to:cyp1a1, cyp1b1, AHRR, IDO1, IDO2, cox2, IL6, VEGFA, cyclinD1, cdc2,MMP-9, c-myc.

Example 4

Synthesis of I-5

Step 1:N-(2-(1H-indol-3-yl)ethyl)-2-chlorothieno[2,3-d]pyrimidin-4-amine

To a solution of 2,4-dichlorothieno[2,3-d]pyrimidine (200 mg, 975.31umol, 1 eq) in i-PrOH (15 mL) was added DIPEA (630.24 mg, 4.88 mmol,849.38 uL, 5.0 eq) and 2-(1H-indol-3-yl)ethanamine (203.14 mg, 1.27mmol, 1.3 eq). The mixture was stirred at 50° C. for 3 h. LC-MS showedthe starting material was consumed completely and one main peak withdesired MS was detected. The reaction mixture was diluted with H₂O (15mL) and extracted with DCM (15 mL×3). The combined organic layers weredried over Na₂SO₄, filtered and concentrated under reduced pressure togive a residue which was purified by flash silica gel chromatography(ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 2030% EtOAc/PEgradient @ 50 mL/min) to give2-chloro-N-[2-(1H-indol-3-yl)ethyl]thieno[2,3-d]pyrimidin-4-amine (260mg, 774.90 umol, 79.4% yield) as a yellow solid. ¹H NMR (400 MHz, CD₃OD)δ ppm 7.67 (d, J=7.9 Hz, 1H), 7.40-7.35 (m, 2H), 7.31 (d, J=8.2 Hz, 1H),7.10-7.05 (m, 2H), 7.00-6.93 (m, 1H), 3.86-3.79 (m, 2H), 3.11 (t, J=7.4Hz, 2H); ES-LCMS m/z 329.0, 331.0 [M+H]+.

Step 2:N-(2-(1H-indol-3-yl)ethyl)-2-(5-fluoropyridin-3-yl)thieno[2,3-d]pyrimidin-4-amine(I-5)

To a solution of2-chloro-N-[2-(1H-indol-3-yl)ethyl]thieno[2,3-d]pyrimidin-4-amine (100mg, 298.04 umol, 1 eq) in 1,4-dioxane (3 mL) and H₂O (1 mL) was added(5-fluoro-3-pyridyl)boronic acid (75.59 mg, 536.47 umol, 1.8 eq), Cs₂CO₃(291.32 mg, 894.11 umol, 3.0 eq) and Pd(dppf)Cl₂ (32.71 mg, 44.71 umol,0.15 eq). The mixture was stirred at 120° C. under microwave for 1 h.LC-MS showed the starting material was consumed completely and one mainpeak with desired MS was detected. The reaction mixture was filtered.The filtrate was diluted with H₂O (15 mL) and extracted with EtOAc (15mL×3). The combined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (HCl condition; column: Phenomenex Kinetex XB-C18150 mm*30 mm, 5 m; mobile phase: [water (0.05% HCl)-ACN]; B %: 50%-75%,12 min) and the desired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]thieno[2,3-d]pyrimidin-4-amine(54.40 mg, 109.05 umol, 36.59% yield, 100% purity, 3HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.19 (s, 1H), 8.84 (s, 1H),8.58 (d, J=9.3 Hz, 1H), 7.63-7.57 (m, 2H), 7.54 (d, J=5.7 Hz, 1H), 7.19(d, J=7.7 Hz, 1H), 7.02-6.90 (m, 3H), 4.08 (t, J=6.8 Hz, 2H), 3.19 (t,J=6.7 Hz, 2H); ES-LCMS m/z 390.0 [M+H]+.

Example 5

Synthesis of I-4

Synthetic Scheme:

Step 1: 2,4-Dichloro-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidine

A mixture of 2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine (1 g, 5.32 mmol, 1eq), 2-bromopropane (3.27 g, 26.59 mmol, 2.50 mL, 5.0 eq) and K₂CO₃(3.68 g, 26.59 mmol, 5 eq) in DMSO (20 mL) was degassed and purged withN₂ for 3 times then the mixture was stirred at 10-20° C. for 48 h underN₂ atmosphere. TLC (PE/EA=3/1, R_(f)=0.50) indicated starting materialwas consumed, and one major new spot with larger polarity was detected.The reaction mixture was diluted with H₂O (50 mL) and extracted withEtOAc (50 mL×3). The combined organic layers were dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 4/1, TLC: PE/EtOAc=3/1, R_(f)=0.50) to give the product2,4-dichloro-7-isopropyl-pyrrolo[2,3-d]pyrimidine (650 mg, 2.71 mmol,50.99% yield, 96% purity) as a white solid. ¹H NMR (400 MHz, CDCl₃) δppm 7.33 (d, J=3.7 Hz, 1H), 6.62 (d, J=3.5 Hz, 1H), 5.20-5.01 (m, 1H),1.53 (d, J=6.8 Hz, 6H); ES-LCMS m/z 230.0, 232.0 [M+H]⁺.

Step 2:2-Chloro-N-[2-(1H-indol-3-yl)ethyl]-7-isopropyl-pyrrolo[2,3-d]pyrimidin-4-amine

A mixture of 2,4-dichloro-7-isopropyl-pyrrolo[2,3-d]pyrimidine (100 mg,417.22 umol, 1 eq), 2-(1H-indol-3-yl)ethanamine (100 mg, 620 umol, 1.2eq) and DIEA (161.77 mg, 1.25 mmol, 218.02 uL, 3 eq) in i-PrOH (5 mL)was degassed and purged with N₂ for 3 times. Then the mixture wasstirred at 50° C. for 11 h under N₂ atmosphere. LC-MS showed 15% ofstarting material was remained. The reaction mixture was concentratedunder reduced pressure to give a residue which was purified on silicagel column chromatography (from PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=3/1,R_(f)=0.28) to give the product2-chloro-N-[2-(1H-indol-3-yl)ethyl]-7-isopropyl-pyrrolo[2,3-d]pyrimidin-4-amine(120 mg, 315.39 umol, 75.59% yield, 93% purity) as light red oil. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.06 (s, 1H), 7.69 (d, J=7.9 Hz, 1H), 7.41 (d,J=7.9 Hz, 1H), 7.26-7.20 (m, 1H), 7.18-7.13 (m, 1H), 7.09 (d, J=2.4 Hz,1H), 6.96 (d, J=3.5 Hz, 1H), 6.23 (s, 1H), 5.22 (s, 1H), 5.03 (d, J=6.8,13.5 Hz, 1H), 3.99 (q, J=6.6 Hz, 2H), 3.17 (t, J=6.6 Hz, 2H), 1.46 (d,J=6.8 Hz, 6H); ES-LCMS m/z 353.8, 354.9 [M+H]⁺.

Step 3:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-7-isopropyl-pyrrolo[2,3-d]pyrimidin-4-amine(I-4)

2-Chloro-N-[2-(1H-indol-3-yl)ethyl]-7-isopropyl-pyrrolo[2,3-d]pyrimidin-4-amine(80 mg, 210.26 umol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (44.44 mg,315.39 umol, 1.5 eq), Pd(dppf)Cl₂ (15.38 mg, 21.03 umol, 0.1 eq) andCs₂CO₃ (205.52 mg, 630.78 umol, 3 eq) in 1,4-dioxane (2 mL) and H₂O (0.5mL) were taken up into a microwave tube. The reaction mixture wasbubbled with N₂ for 3 min then sealed and heated at 110° C. for 30 minunder microwave. LC-MS showed 90% of desired compound was detected. Thereaction mixture was concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (MeCN/H₂O as eluents,acidic condition, Instrument: DC/Phenomenex Kinetex XB-C18 150 mm*30 mm,5 μm/Mobile phase: water (0.05% HCl)-ACN/Gradient: B from 47% to 77% in10 min/Flow rate: 25 mL/min) followed by lyophilization to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-7-isopropyl-pyrrolo[2,3-d]pyrimidin-4-amine(51.57 mg, 120.95 umol, 57.52% yield, 97.21% purity, 3HCl salt) as ayellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.41 (s, 1H), 8.72 (s,1H), 8.50 (d, J=8.6 Hz, 1H), 8.09 (s, 1H), 7.64 (d, J=7.7 Hz, 1H), 7.40(s, 1H), 7.35 (d, J=8.2 Hz, 1H), 7.23 (s, 1H), 7.07 (t, J=7.4 Hz, 1H),7.02-6.93 (m, 1H), 6.69 (s, 1H), 5.06 (s, 1H), 3.89 (s, 1H), 3.80-3.77(m, 2H), 3.20-3.00 (m, 2H), 1.48 (d, J=6.6 Hz, 6H); ES-LCMS m/z 414.9[M+H]⁺.

Example 6

Synthesis of I-3

Synthetic Scheme:

Step 1:N-(2-(1H-indol-3-yl)ethyl)-7-bromo-2-chlorothieno[3,2-d]pyrimidin-4-amine

To a solution of 7-bromo-2,4-dichloro-thieno[3,2-d]pyrimidine (100 mg,352.16 umol, 1 eq) and 2-(1H-indol-3-yl)ethanamine (84.63 mg, 528.24umol, 1.5 eq) in i-PrOH (3 mL) was added DIEA (136.54 mg, 1.06 mmol,184.02 uL, 3 eq). The mixture was stirred at 50° C. for 3 h. LC-MSshowed no starting material was remained and 93% of desired compound wasdetected. The reaction mixture was concentrated under reduced pressureto give a residue which was purified on silica gel column chromatography(from DCM/MeOH=1/0 to 100/1, TLC: PE/EtOAc=3/1, R_(f)=0.42) to give7-bromo-2-chloro-N-[2-(1H-indol-3-yl)ethyl]thieno[3,2-d]pyrimidin-4-amine(140 mg, 339.94 umol, 96.53% yield, 99% purity) as gray solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.11 (s, 1H), 7.69-7.66 (m, 2H), 7.43 (d, J=8.2Hz, 1H), 7.25-7.23 (m, 1H), 7.18-7.14 (m, 1H), 7.09 (d, J=2.2 Hz, 1H),5.25 (s, 1H), 4.01 (q, J=6.4 Hz, 2H), 3.18 (t, J=6.5 Hz, 2H); ES-LCMSm/z 406.9, 408.9 [M+H]⁺.

Step 2:N-(2-(1H-indol-3-yl)ethyl)-2-chloro-7-(prop-1-en-2-yl)thieno[3,2-d]pyrimidin-4-amine

7-Bromo-2-chloro-N-[2-(1H-indol-3-yl)ethyl]thieno[3,2-d]pyrimidin-4-amine(120 mg, 291.38 umol, 1 eq),2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (39.17 mg, 233.10umol, 0.8 eq), Pd(dppf)Cl₂ (42.64 mg, 58.28 umol, 0.2 eq) and Cs₂CO₃(284.81 mg, 874.14 umol, 3.0 eq) in 1,4-dioxane (5 mL) and H₂O (1 mL)were taken up into a microwave tube. The tube was purged with N₂ for 3min then sealed and heated at 80° C. for 10 min under microwave. LC-MSshowed most of starting material was consumed and 68% of desiredcompound was detected. The combined reaction mixture was diluted withH₂O (20 mL) and extracted with EtOAc (20 mL×3). The combined organiclayers were dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified on silica gelcolumn chromatography (from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1,R_(f)=0.45) to give the productN-(2-(1H-indol-3-yl)ethyl)-2-chloro-7-(prop-1-en-2-yl)thieno[3,2-d]pyrimidin-4-amine(95 mg, 73.85% yield, 98% purity) as light yellow oil; ¹H NMR (400 MHz,CDCl₃) δ ppm 8.10 (s, 1H), 7.69 (d, J=7.8 Hz, 1H), 7.51 (s, 1H), 7.42(d, J=8.0 Hz, 1H), 7.26-7.22 (m, 1H), 7.20-7.13 (m, 1H), 7.09 (d, J=2.3Hz, 1H), 6.37 (s, 1H), 5.38 (s, 1H), 5.11 (s, 1H), 4.00 (q, J=6.4 Hz,2H), 3.18 (t, J=6.7 Hz, 2H), 2.21 (s, 3H); ES-LCMS m/z 369.0, 371.0[M+H]⁺.

Step 3:N-(2-(1H-indol-3-yl)ethyl)-2-(5-fluoropyridin-3-yl)-7-(prop-1-en-2-yl)thieno[3,2-d]pyrimidin-4-amine

2-Chloro-N-[2-(1H-indol-3-yl)ethyl]-7-isopropenyl-thieno[3,2-d]pyrimidin-4-amine(95 mg, 252.38 umol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (53.34 mg,378.58 umol, 1.5 eq), Pd(dppf)Cl₂ (18.47 mg, 25.24 umol, 0.1 eq) andCs₂CO₃ (246.69 mg, 757.15 umol, 3 eq) in 1,4-dioxane (2 mL) and H₂O (0.5mL) were taken up into a microwave tube. The sealed tube was purged withN₂ for 3 min then heated at 110° C. for 30 min under microwave. LC-MSshowed most of starting material was consumed and 85% of desiredcompound was detected. The reaction mixture was concentrated underreduced pressure to give a residue which was purified on silica gelcolumn chromatography (from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1,R_(f)=0.44) to give the product2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-7-isopropenyl-thieno[3,2-d]pyrimidin-4-amine(85 mg, 188.00 umol, 74.49% yield, 95% purity) as oil. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.59 (s, 1H), 8.55 (d, J=2.6 Hz, 1H), 8.48 (d, J=10.1 Hz,1H), 8.07 (s, 1H), 7.69 (d, J=7.9 Hz, 1H), 7.56 (s, 1H), 7.42 (d, J=7.9Hz, 1H), 7.26-7.22 (m, 1H), 7.20-7.15 (m, 1H), 7.11 (s, 1H), 6.70 (s,1H), 5.46 (s, 1H), 5.06 (s, 1H), 4.14 (q, J=6.5 Hz, 2H), 3.26 (t, J=6.5Hz, 2H), 2.30 (s, 3H); ES-LCMS m/z 430.0 [M+H]⁺.

Step 4:N-(2-(1H-indol-3-yl)ethyl)-2-(5-fluoropyridin-3-yl)-7-isopropylthieno[3,2-d]pyrimidin-4-amine(I-3)

To a solution of2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-7-isopropenyl-thieno[3,2-d]pyrimidin-4-amine(50 mg, 110.59 umol, 1 eq) in MeOH (20 mL) and THE (2 mL) was added Pd/C(10%, 50 mg) under N₂. The suspension was degassed under vacuum andpurged with H₂ for 3 times. The mixture was stirred under H₂ (15 psi) at10-15° C. for 0.5 h. LC-MS showed no starting material was remained and95% of desired compound was detected. The reaction mixture was filteredand the filtrate was concentrated under reduced pressure. The residuewas purified by preparative HPLC (MeCN/H₂O as eluents, acidic condition,Instrument: DC/Phenomenex Kinetex XB-C18 150 mm*30 mm, 5 μm/Mobilephase: water (0.05% HC)-ACN/Gradient: B from 62% to 92% in 10 min/Flowrate: 25 mL/min) followed by lyophilization to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-7-isopropyl-thieno[3,2-d]pyrimidin-4-amine(40.32 mg, 74.05 umol, 66.96% yield, 99.34% purity, 3HCl salt) as ayellow solid. 1H NMR (400 MHz, DMSO-d₆) δ ppm 10.85 (s, 1H), 9.46 (s,1H), 8.69 (d, J=2.8 Hz, 1H), 8.42 (d, J=10.0 Hz, 1H), 8.19 (t, J=5.6 Hz,1H), 7.77 (s, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.34 (d, J=8.0 Hz, 1H), 7.22(d, J=2.0 Hz, 1H), 7.07 (t, J=7.4 Hz, 1H), 6.99-6.95 (m, 1H), 3.93-3.87(m, 2H), 3.46-3.41 (m, 1H), 3.12 (t, J=7.4 Hz, 2H), 1.37 (d, J=6.8 Hz,6H); ES-LCMS m/z 432.0 [M+H]⁺.

Example 7

Synthesis of I-2

Synthetic Scheme:

Step 1: 2-Formyl-3-methylbutanenitrile

To a mixture of diisopropylamine (2.43 g, 24.06 mmol, 3.40 mL, 1 eq) inTH (20 mL) was added n-BuLi (2.5 M, 10.10 mL, 1.05 eq) dropwise at −78°C. under N₂. The mixture was stirred at −78° C. for 10 min, then warmedup to 0° C. and stirred for 1 h. The mixture was cooled to −78° C.,3-methylbutanenitrile (2 g, 24.06 mmol, 2.53 mL, 1 eq) dissolved in THE(15 mL, anhydrous) was added dropwise and stirred at −78° C. for 10 min.A solution of ethyl formate (1.87 g, 25.26 mmol, 2.03 mL, 1.05 eq) inTHE (15 mL, anhydrous) was added dropwise and stirred at −78° C. for 40min, then the mixture was warmed to 5-14° C. for 16 h. TLC (PE/EA=3/1,R_(f)=0.34) indicated one major new spot was detected. The reactionmixture was quenched by addition 1 N HCl solution (50 mL) at −78° C.,and extracted with EtOAc (50 mL×3). The combined organic layers weredried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to give a residue which was purified on silica gel columnchromatography (from PE/EtOAc=1/0 to 1/4, TLC: PE/EtOAc=3/1, R_(f)=0.34)to give 2-formyl-3-methyl-butanenitrile (2 g, 16.20 mmol, 67.32% yield,90% purity) as light yellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.57 (s,1H), 3.43 (d, J=4.4 Hz, 1H), 2.56-2.40 (m, 1H), 1.20 (d, J=6.8 Hz, 3H),1.12 (d, J=6.8 Hz, 3H).

Step 2: 4-Isopropyl-1H-pyrazol-5-amine

A mixture of 2-formyl-3-methyl-butanenitrile (500 mg, 4.05 mmol, 1 eq),hydrazine hydrate (168.67 mg, 5.26 mmol, 190.37 uL, 1.3 eq) and AcOH(425.50 mg, 7.09 mmol, 405.24 uL, 1.75 eq) in EtOH (20 mL) was degassedand purged with N₂ for 3 times. The mixture was stirred at 80-90° C.(reflux) for 16 h under N₂ atmosphere then concentrated under reducedpressure. The residue was diluted with sat. NaHCO₃ solution (20 mL) andextracted with DCM (20 mL×3). The combined organic layers were driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto give the crude product 4-isopropyl-1H-pyrazol-5-amine (420 mg, 3.02mmol, 74.58% yield, 90% purity) as yellow solid, which was used in thenext step without further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm7.13 (s, 1H), 2.69 (t, J=6.9 Hz, 1H), 1.21 (d, J=6.8 Hz, 6H).

Step 3: Methyl 3-(5-fluoropyridin-3-yl)-3-oxopropanoate

To a solution of 5-fluoropyridine-3-carboxylic acid (500 mg, 3.54 mmol,1 eq) in THE (20 mL) was added CDI (689.51 mg, 4.25 mmol, 1.2 eq). Themixture was stirred at 5-14° C. for 2 h.(3-Methoxy-3-oxo-propanoyl)oxypotassium (553.43 mg, 3.54 mmol, 1 eq) andMgCl₂ (337.39 mg, 3.54 mmol, 1 eq) was added and the reaction wasstirred at 5-14° C. for 16 h. TLC (PE/EA=1/1, R_(f)=0.58) indicated thestarting material was consumed and one major new spot with largerpolarity was detected. The reaction mixture was diluted with 3 N HClsolution (20 mL) and extracted with EtOAc (20 mL×3). The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified on silica gel column chromatography (from PE/EtOAc=1/0 to 1/4,TLC: PE/EtOAc=1/1, R_(f)=0.58) to give methyl3-(5-fluoro-3-pyridyl)-3-oxo-propanoate (175 mg, 843.21 umol, 23.80%yield, 95% purity) as a white solid and methyl3-(5-fluoro-3-pyridyl)-3-oxo-propanoate (175 mg, 843.21 umol, 23.80%yield, 95% purity) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 12.48(s, 1H), 8.97 (s, 1H), 8.82 (s, 1H), 8.70 (d, J=2.6 Hz, 1H), 8.56 (d,J=2.6 Hz, 1H), 7.96 (d, J=2.2, 8.6 Hz, 1H), 7.86-7.73 (m, 1H), 5.75 (s,1H), 4.04 (s, 2H), 3.84 (s, 3H), 3.78 (s, 3H); ES-LCMS m/z 198.1 [M+H]⁺.

Step 4:5-(5-Fluoropyridin-3-yl)-3-isopropylpyrazolo[1,5-a]pyrimidin-7-ol

To a solution of methyl 3-(5-fluoro-3-pyridyl)-3-oxo-propanoate (200 mg,963.67 umol, 1 eq) in AcOH (5 mL) was added4-isopropyl-1H-pyrazol-5-amine (134.03 mg, 963.67 umol, 1 eq). Themixture was stirred at 120° C. for 0.5 h. LCMS showed starting materialwas consumed completely and one main peak with desired MS was detected.The reaction mixture was concentrated under reduced pressure to give thecrude product5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-ol (300mg, 639.05 umol, 66.31% yield, 58% purity) as yellow oil, which was usedin the next step without further purification. ¹H NMR (400 MHz, CD₃OD) δppm 8.79 (m, 1H), 8.63-8.67 (m, 1H), 8.09-8.06 (m, 1H), 7.87 (m, 1H),6.06 (m, 1H), 3.26-3.25 (m, 1H), 2.75-2.70 (m, 1H), 1.30 (d, J=6.8 Hz,3H), 1.15-1.09 (m, 3H); ES-LCMS m/z 567.0 [2M+Na]⁺.

Step 5:7-Chloro-5-(5-fluoropyridin-3-yl)-3-isopropylpyrazolo[1,5-a]pyrimidine

A solution of5-(5-fluoropyridin-3-yl)-3-isopropylpyrazolo[1,5-a]pyrimidin-7-ol (300mg, 639.05 umol, 1 eq) in POCl₃ (4.95 g, 32.28 mmol, 3 mL, 50.52 eq) wasstirred at 110° C. for 3 h. LC-MS showed no starting material wasremained and 67% of desired compound was detected. The reaction mixturewas concentrated under reduced pressure then diluted with DCM (20 mL×2),concentrated under reduced pressure to give a residue which was purifiedon silica gel column chromatography (from PE/EtOAc=1/0 to 2/5, TLC:PE/EtOAc=1/1, R_(f)=0.64) to give the product7-chloro-5-(5-fluoropyridin-3-yl)-3-isopropylpyrazolo[1,5-a]pyrimidine(140 mg, 385.25 umol, 60.28% yield, 80% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.10 (s, 1H), 8.61-8.60 (m, 1H), 8.22-8.14(m, 2H), 7.41-7.27 (m, 1H), 3.47-3.38 (m, 1H), 1.48-1.44 (m, 6H);ES-LCMS m/z 291.0, 293.0[M+H]⁺.

Step 6:N-(2-(1H-indol-3-yl)ethyl)-5-(5-fluoropyridin-3-yl)-3-isopropylpyrazolo[1,5-a]pyrimidin-7-amine(I-2)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(80 mg, 220.14 umol, 1 eq) and 2-(1H-indol-3-yl)ethanamine (52.90 mg,330.21 umol, 1.5 eq) in i-PrOH (3 mL) was added DIEA (85.36 mg, 660.42umol, 115.03 uL, 3.0 eq). The mixture was stirred at 50° C. for 3 h.LCMS showed no starting material was remained and 92% of desiredcompound was detected. The reaction mixture was concentrated underreduced pressure and the residue was purified by preparative HPLC(MeCN/H₂O as eluents, acidic condition, Instrument: DC/PhenomenexKinetex XB-C18 150 mm*30 mm, 5 μm/Mobile phase: water (0.05%HCl)-ACN/Gradient: B from 47% to 57% in 10 min/Flow rate: 25 mL/min)followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(88.45 mg, 168.66 umol, 76.61% yield, 99.89% purity, 3HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.73 (d, J=2.2 Hz, 1H), 8.34(s, 1H), 8.15 (s, 1H), 7.55 (d, J=8.6 Hz, 1H), 7.43 (d, J=7.9 Hz, 1H),7.07 (d, J=8.2 Hz, 1H), 6.94 (s, 1H), 6.86 (t, J=7.2 Hz, 1H), 6.72 (t,J=7.4 Hz, 1H), 5.71 (s, 1H), 3.95 (t, J=5.7 Hz, 2H), 3.17-3.12 (m, 3H),1.26 (d, J=6.8 Hz, 6H); ES-LCMS m/z 415.2 [M+H]⁺.

Example 8

Synthesis of I-10

Synthetic Scheme:

Step 1: tert-Butyl N-[2-(2-nitroanilino)ethyl]carbamate

To a solution of 1-fluoro-2-nitro-benzene (10 g, 70.87 mmol, 7.46 mL, 1eq) and tert-butyl N-(2-aminoethyl)carbamate (11.35 g, 70.87 mmol, 11.13mL, 1 eq) in DMF (50 mL) was added K₂CO₃ (15.67 g, 113.39 mmol, 1.6 eq).The mixture was stirred at 70° C. for 18 h. TLC (PE/EtOAc=5/1,R_(f)=0.69) showed most starting material was consumed. The mixture wasdiluted with H₂O (100 mL), extracted with EtOAc (100 mL×3). The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure. The crude material was purified onsilica gel column chromatography (from PE/EtOAc=1/0 to 5/4, TLC:PE/EtOAc=5/1, R_(f)=0.69) to give tert-butylN-[2-(2-nitroanilino)ethyl]carbamate (19 g, 67.54 mmol, 95.3% yield,100% purity) as an orange solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.19 (d,J=8.8 Hz, 1H), 7.47-7.43 (m, 1H), 6.95 (d, J=8.4 Hz, 1H), 6.69-6.65 (m,1H), 3.50-3.43 (m, 4H), 1.46 (s, 9H); ES-LCMS m/z 304.0 [M+Na]⁺.

Step 2: tert-Butyl N-[2-(2-aminoanilino)ethyl]carbamate

To a solution of tert-butyl N-[2-(2-nitroanilino)ethyl]carbamate (5 g,17.77 mmol, 1 eq) in MeOH (50 mL) was added Pd/C (10%, 500 mg) under N₂atmosphere. The suspension was degassed and purged with H₂ for 3 times.The mixture was stirred under H₂ (15 Psi) at 2-9° C. for 18 h. LC-MSshowed the staring material was consumed completely and one main peakwith desired MS was detected. The mixture was filtered and concentratedunder reduced pressure to give crude tert-butylN-[2-(2-aminoanilino)ethyl]carbamate (4.4 g, 17.51 mmol, 98.50% yield,crude) as dark red oil which was used in the next step without furtherpurification. ¹H NMR (400 MHz, CD₃OD) δ ppm 6.70-6.67 (m, 2H), 6.62 (d,J=7.6 Hz, 1H), 6.56 (t, J=7.2 Hz, 1H), 3.29-3.26 (m, 2H), 3.18-3.17 (m,2H), 1.42 (s, 9H); ES-LCMS m/z 252.2 [M+H]⁺.

Step 3: tert-Butyl N-[2-(2-oxo-3H-benzimidazol-1-yl)ethyl]carbamate

To a solution of tert-butyl N-[2-(2-aminoanilino)ethyl]carbamate (2 g,7.96 mmol, 1 eq) in THF (20 mL) was added CDI (1.55 g, 9.55 mmol, 1.2eq). The mixture was stirred at 80° C. for 2 h. LC-MS showed startingmaterial was consumed completely and one main peak with desired MS wasdetected. The mixture was diluted with EtOAc (50 mL), washed with citricacid solution (aq., 20 mL×2), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedon silica gel column chromatography (from PE/EtOAc=1/0 to 5/4, TLC:PE/EtOAc=1/1, R_(f)=0.31). The slight yellow oil obtained was dilutedwith 1 N HCl solution (5 mL), filtered and collected the solid. Thesolid was triturated with isopropyl ether (20 mL), filtered andcollected the product tert-butyl N-[2-(2-oxo-3H-benzimidazol-1-yl)ethyl]carbamate (1.63 g, 5.73 mmol, 72.0% yield, 97.52% purity) as a whitesolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 7.13-7.01 (m, 4H), 3.93 (t, J=6.0Hz, 2H), 3.35-3.31 (m, 2H), 1.29, 1.17 (s, 9H); ES-LCMS m/z 300.1[M+H]⁺.

Step 4: 3-(2-Aminoethyl)-1H-benzimidazol-2-one

To a solution of tert-butylN-[2-(2-oxo-3H-benzimidazol-1-yl)ethyl]carbamate (646.02 mg, 2.27 mmol,1 eq) in DCM (10 mL) was added HCl/1,4-dioxane (4 M, 5.00 mL, 8.80 eq)dropwise at 0° C. After addition, the mixture was stirred at 4-12° C.for 1 h. LC-MS showed most of starting material was consumed. Thereaction mixture was concentrated under reduced pressure to give crude3-(2-aminoethyl)-1H-benzimidazol-2-one (400 mg, 1.87 mmol, 82.4% yield,HCl salt) as a white solid, which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, CD₃OD) δ ppm 7.18-7.17 (m, 1H),7.11-7.08 (m, 3H), 4.19-4.16 (m, 2H), 3.31-3.28 (m, 2H); ES-LCMS m/z178.1 [M+H]⁺.

Step 5:3-[2-[(2-Chloro-9-isopropyl-purin-6-yl)amino]ethyl]-1H-benzimidazol-2-one

To a solution of 2,6-dichloro-9-isopropyl-purine (200 mg, 827.24 umol, 1eq) in isopropanol (10 mL) was added DIEA (427.65 mg, 3.31 mmol, 576.35uL, 4.0 eq) and 3-(2-aminoethyl)-1H-benzimidazol-2-one (265.13 mg, 1.24mmol, 1.5 eq, HCl). The mixture was stirred at 50° C. for 5 h. LC-MSshowed most of starting material was consumed. The reaction mixture wasconcentrated under reduced pressure to remove isopropanol. The residuewas diluted with H₂O (20 mL) and extracted with DCM/isopropanol (20mL×3, v/v=3/1). The combined organic layers were dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was purified on silica gel column chromatography(from DCM/MeOH=1/0 to 10/1, TLC: DCM/MeOH=10/1, R_(f)=0.15) to give3-[2-[(2-chloro-9-isopropyl-purin-6-yl)amino]ethyl]-1H-benzimidazol-2-one(280 mg, 730.45 umol, 88.3% yield, 97% purity) as a white solid; ¹H NMR(400 MHz, CD₃OD) δ ppm 8.14 (s, 1H), 7.28 (d, J=7.2 Hz, 1H), 7.06-6.97(m, 3H), 4.78-4.75 (m, 1H), 4.22 (t, J=6.0 Hz, 2H), 3.97 (t, J=6.0 Hz,2H), 1.62 (d, J=6.4 Hz, 6H); ES-LCMS m/z 372.1, 374.0 [M+H]⁺.

Step 6:3-[2-[[2-(5-Fluoro-3-pyridyl)-9-isopropyl-purin-6-yl]amino]ethyl]-1H-benzimidazol-2-one(I-10)

To a solution of3-[2-[(2-chloro-9-isopropyl-purin-6-yl)amino]ethyl]-1H-benzimidazol-2-one(100 mg, 260.88 umol, 1 eq) and (5-fluoro-3-pyridyl)boronic acid (73.52mg, 521.75 umol, 2 eq) in 1,4-dioxane (2 mL) and H₂O (0.5 mL) was addedPd(dppf)Cl₂ (9.54 mg, 13.04 umol, 0.05 eq) and Cs₂CO₃ (255.00 mg, 782.63umol, 3 eq). The mixture was purged with N₂ for 3 min and stirred at120° C. for 0.5 h under microwave. LC-MS showed most of startingmaterial was consumed and desired compound was detected. The reactionmixture was concentrated under reduced pressure. The residue was dilutedwith EtOAc/MeOH/THF (10 mL/10 mL/10 mL) and stirred for 5 min, filtered.The filtrate was concentrated under reduced pressure to give a residuewhich was purified by preparative HPLC (MeCN/H₂O as eluents, acidiccondition, Instrument: DC/Phenomenex Kinetex XB-C18 150 mm*30 mm, 5μm/Mobile phase: water (0.05% HCl)-ACN/Gradient: B from 20% to 50% in 12min/Flow rate: 25 mL/min) followed by lyophilization to yield3-[2-[[2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-yl]amino]ethyl]-1H-benzimidazol-2-one(11.08 mg, 21.29 umol, 8.16% yield, 97.1% purity, 2HCl salt, 10.85 mgwas delivered) as a light yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm9.51-9.27 (m, 2H), 9.15-8.90 (m, 2H), 7.23 (d, J=7.8 Hz, 1H), 6.92 (t,J=7.8 Hz, 1H), 6.75 (t, J=7.5 Hz, 1H), 6.63 (d, J=7.8 Hz, 1H), 5.19-4.97(m, 1H), 4.30-4.15 (m, 4H), 1.67 (d, J=6.8 Hz, 6H); ES-LCMS m/z 433.1[M+H]⁺.

Example 9

Synthesis of I-19

Synthetic Scheme:

Step 1:N-(2-(1H-Indol-3-yl)ethyl)-7-bromo-2-chloropyrrolo[2,1-f][1,2,4]triazin-4-amine

To a solution of 7-bromo-2,4-dichloro-pyrrolo[2,1-f][1,2,4]triazine (100mg, 374.66 umol, 1 eq) in i-PrOH (3 mL) was added DIEA (242.10 mg, 1.87mmol, 326.29 uL, 5.0 eq) and 2-(1H-indol-3-yl)ethanamine (78.03 mg,487.06 umol, 1.3 eq). The mixture was stirred at 50° C. for 1.5 h. LC-MSshowed the starting material was consumed completely and one main peakwith desired MS was detected. The reaction mixture was diluted with H₂O(15 mL) and extracted with DCM (15 mL×3). The combined organic layerswere dried over Na₂SO₄, filtered and concentrated under reduced pressureto give a residue which was purified by flash silica gel chromatography(ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 2025% Ethylacetate/Petroleum ether gradient @55 mL/min) to give7-bromo-2-chloro-N-[2-(1H-indol-3-yl)ethyl]pyrrolo[2,1-f][1,2,4]triazin-4-amine(140 mg, 354.78 umol, 94.7% yield) as a white solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 8.08 (s, 1H), 7.66 (d, J=7.7 Hz, 1H), 7.42 (d, J=8.2 Hz,1H), 7.26-7.23 (m, 1H), 7.19-7.14 (m, 1H), 7.09 (d, J=2.2 Hz, 1H), 6.59(d, J=4.6 Hz, 1H), 6.40 (br s, 1H), 5.55 (m, 1H), 3.99 (q, J=6.2 Hz,2H), 3.17 (t, J=6.6 Hz, 2H); ES-LCMS m/z 389.9, 392.0 [M+H]⁺.

Step 2:N-(2-(1H-Indol-3-yl)ethyl)-2-chloro-7-(prop-1-en-2-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

To a solution of7-bromo-2-chloro-N-[2-(1H-indol-3-yl)ethyl]pyrrolo[2,1-f][1,2,4]triazin-4-amine(90 mg, 228.07 umol, 1 eq) in 1,4-dioxane (2 mL) and H₂O (0.5 mL) wasadded 2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (38.33 mg,228.07 umol, 1.0 eq), Pd(dppf)Cl₂ (33.38 mg, 45.61 umol, 0.2 eq) andCs₂CO₃ (222.93 mg, 684.22 umol, 3.0 eq). The mixture was stirred undermicrowave at 80° C. for 30 min. LCMS showed 67% of desired product wasfound. The reaction mixture was filtered and the filtrate was dilutedwith H₂O (15 mL) then extracted with EtOAc (15 mL×3). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by flash silicagel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluentof 10˜13% Ethyl acetate/Petroleum ether gradient @ 50 mL/min) to give2-chloro-N-[2-(1H-indol-3-yl)ethyl]-7-isopropenyl-pyrrolo[2,1-f][1,2,4]triazin-4-amine(65 mg, 144.10 umol, 63.2% yield) as colorless oil. ¹H NMR (400 MHz,CDCl₃) δ ppm 8.08 (br s, 1H), 7.67 (d, J=7.7 Hz, 1H), 7.41 (d, J=8.2 Hz,1H), 7.26-7.23 (m, 1H), 7.20-7.14 (m, 1H), 7.09 (d, J=2.2 Hz, 1H), 6.58(d, J=4.6 Hz, 1H), 6.36 (br s, 1H), 6.25 (s, 1H), 5.50 (br s, 1H), 5.35(s, 1H), 4.02-3.96 (m, 2H), 3.16 (t, J=6.5 Hz, 2H), 2.20 (s, 3H);ES-LCMS m/z 352.0, 354.0 [M+H]⁺.

Step 3:N-(2-(1H-Indol-3-yl)ethyl)-2-(5-fluoropyridin-3-yl)-7-(prop-1-en-2-yl)pyrrolo[2,1-f][1,2,4]triazin-4-amine

To a solution of2-chloro-N-[2-(1H-indol-3-yl)ethyl]-7-isopropenyl-pyrrolo[2,1-f][1,2,4]triazin-4-amine(65 mg, 144.10 umol, 1 eq) in 1,4-dioxane (3 mL) and H₂O (1 mL) wasadded (5-fluoro-3-pyridyl)boronic acid (30.46 mg, 216.15 umol, 1.5 eq),Pd(dppf)Cl₂ (15.82 mg, 21.62 umol, 0.15 eq) and Cs₂CO₃ (140.85 mg,432.31 umol, 3.0 eq). The mixture was stirred at 110° C. under microwavefor 0.5 h. LCMS showed about 72% of desired product was detected. Thereaction mixture was filtered and the filtrate was diluted with H₂O (15mL) then extracted with EtOAc (15 mL×3). The combined organic layerswere dried over Na₂SO₄, filtered and concentrated under reduced pressureto give a residue which was purified by preparative TLC (PE/EtOAc=1.5/1,R_(f)=0.6) to give2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-7-isopropenyl-pyrrolo[2,1-f][1,2,4]triazin-4-amine(40 mg, 76.61 umol, 53.2% yield) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.29 (s, 1H), 8.38 (d, J=2.9 Hz, 1H), 8.23-8.16 (m, 1H),8.05 (br s, 1H), 7.55 (d, J=7.7 Hz, 1H), 7.30-7.27 (m, 1H), 7.30-7.24(m, 1H), 7.14-7.09 (m, 1H), 7.07-7.02 (m, 1H), 6.95 (d, J=2.2 Hz, 1H),6.54 (d, J=4.6 Hz, 1H), 6.34-6.23 (m, 2H), 5.45 (t, J=5.6 Hz, 1H), 5.27(s, 1H), 3.95 (q, J=6.5 Hz, 2H), 3.09 (t, J=6.6 Hz, 2H), 2.15 (s, 3H);ES-LCMS m/z 413.1 [M+H]⁺.

Step 4:N-(2-(1H-Indol-3-yl)ethyl)-2-(5-fluoropyridin-3-yl)-7-isopropylpyrrolo[2,1-f][1,2,4]triazin-4-amine(I-19)

To a solution of2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-7-isopropenyl-pyrrolo[2,1-f][1,2,4]triazin-4-amine(30 mg, 57.46 umol, 1 eq) in MeOH (8 mL) was added Pd/C (10%, 40 mg).The suspension was degassed under vacuum and purged with H₂ severaltimes then the mixture was stirred at H₂ (15 psi) at 20° C. for 0.5 h.LC-MS showed the starting material was consumed completely and one mainpeak with desired MS was detected. The reaction mixture was filtered andconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (HCl condition; column: Phenomenex Kinetex XB-C18150 mm*30 mm, 5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 75%-95%,12 min) and the desired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-7-isopropyl-pyrrolo[2,1-f][1,2,4]triazin-4-amine(6.75 mg, 12.89 umol, 17.72% yield, 100% purity, 3HCl salt) as a whitesolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.05 (s, 1H), 8.76 (br s, 1H), 8.42(d, J=8.8 Hz, 1H), 7.63 (d, J=7.3 Hz, 1H), 7.23 (d, J=7.7 Hz, 1H),7.06-6.97 (m, 3H), 6.92 (d, J=4.0 Hz, 1H), 6.54 (d, J=4.0 Hz, 1H), 4.01(t, J=7.1 Hz, 2H), 3.55 (d, J=7.0, 14.1 Hz, 1H), 3.17 (t, J=6.8 Hz, 2H);ES-LCMS m/z 415.1 [M+H]⁺.

Example 10

Synthesis of I-22

Step 1: 2-(Chloromethyl)imidazo[1,2-a]pyridine

To a solution of pyridin-2-amine (5 g, 53.13 mmol, 1 eq) in DME (10 mL)was added 1,3-dichloropropan-2-one (13.49 g, 106.25 mmol, 2 eq). Themixture was stirred at 20° C. for 1 h. The reaction mixture wasconcentrated under reduced pressure. The residue was dissolved in EtOH(50 mL) then stirred at 90° C. for 16 h. LC-MS showed the startingmaterial was consumed completely and desired MS was detected. Thereaction mixture was concentrated and the residue was hydrolyzed withsaturated solution of sodium carbonate (100 mL). The reaction mixturewas extracted with DCM (60 mL×3), combined, dried with Na₂SO₄, filtered,and concentrated under reduced pressure to yield2-(chloromethyl)imidazo[1,2-a]pyridine (8.85 g, crude), which was usedin the next step without further purification. ¹H NMR (400 MHz, CDCl₃) δppm 8.14 (d, J=6.8 Hz, 1H), 7.70-7.59 (m, 2H), 7.26-7.20 (m, 1H), 6.85(t, J=6.8 Hz, 1H), 4.79 (s, 2H); ES-LCMS m/z 167.2, 169.5 [M+H]⁺.

Step 2: 2-Imidazo[1,2-a]pyridin-2-ylacetonitrile

A mixture of 2-(chloromethyl)imidazo[1,2-a]pyridine (3 g, 18.01 mmol, 1eq) and KCN (1.52 g, 23.41 mmol, 1.3 eq) in DMSO (50 mL) was degassedand purged with N₂ for 3 times. Then the mixture was stirred at 80° C.for 16 h under N₂ atmosphere. LC-MS showed 27% of the starting materialwas remained and 17% of desired compound was detected. The reactionmixture was quenched with water (200 mL) then extracted with EtOAc (100mL×3). The combined organic layers were washed with brine (50 mL), driedover Na₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromDCM/MeOH=1/0 to 10/1, TLC: DCM/MeOH=10/1, R_(f)=0.49) to yield2-imidazo[1,2-a]pyridin-2-ylacetonitrile (200 mg, 890.75 umol, 5.0%yield, 70% purity) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm8.59-8.49 (m, 1H), 7.92 (s, 1H), 7.55-7.49 (m, 1H), 7.28-7.11 (m, 1H),6.92-6.85 (m, 1H), 4.12 (s, 3H); ES-LCMS m/z 158.1 [M+H]⁺.

Step 3: 2-Imidazo[1,2-a]pyridin-2-ylethanamine

To a solution of 2-imidazo[1,2-a]pyridin-2-ylacetonitrile (100 mg,445.37 umol, 1 eq) in THE (5 mL) was added BH₃-Me₂S (10 M, 445.37 uL, 10eq). The mixture was stirred at 80° C. for 2 h until LC-MS showed thestarting material was consumed completely and desired MS was detected.The reaction mixture was quenched with MeOH at 10° C. followed bystirring at 80° C. for 1 h. The resulting mixture was concentrated underreduced pressure to yield 2-imidazo[1,2-a]pyridin-2-ylethanamine (71.8mg, crude) as a white solid, which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.82-8.61 (m, 1H),8.21-8.06 (m, 1H), 8.02-7.61 (m, 2H), 7.52-7.21 (m, 1H), 3.61-3.38 (m,2H), 3.35-3.28 (m, 2H); ES-LCMS m/z 162.1 [M+H]⁺.

Step 4:2-(5-Fluoro-3-pyridyl)-N-(2-imidazo[1,2-a]pyridin-2-ylethyl)-9-isopropyl-purin-6-amine(I-22)

To a solution of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine (50mg, 164.55 umol, 1 eq) in i-PrOH (5 mL) was added DIEA (106.33 mg,822.73 umol, 143.31 uL, 5 eq) and 2-imidazo[1,2-a]pyridin-2-ylethanamine(71.8 mg, 445.40 umol, 2.71 eq). The mixture was stirred at 90° C. for16 h. LC-MS showed the starting material was consumed completely and onemain peak with desired MS was detected. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Kinetex XB-C18 150 mm×30 mm, 5m; mobile phase: [water (0.05% HCl)-ACN]; B %: 10%-38%, 12 min) and thedesired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N-(2-imidazo[1,2-a]pyridin-2-ylethyl)-9-isopropyl-purin-6-amine(22.38 mg, 41.71 umol, 25.4% yield, 98% purity, 3HCl) as a yellow solid.¹H NMR (400 MHz, CD₃OD) δ ppm 9.62 (s, 1H), 9.36 (s, 1H), 9.18-8.90 (m,2H), 8.71 (d, J=6.2 Hz, 1H), 8.17 (s, 1H), 7.96-7.76 (m, 2H), 7.43 (t,J=6.8 Hz, 1H), 5.16 (s, 1H), 4.30 (s, 2H), 3.43 (t, J=6.4 Hz, 2H), 1.75(d, J=6.8 Hz, 6H); ES-LCMS m/z 417.1 [M+H]⁺.

Example 11

Synthesis of I-20

Synthetic Scheme:

Step 1: 4,6-Dichloro-2-iodo-pyrimidine

To a solution of 4,6-dichloropyrimidin-2-amine (5 g, 30.49 mmol, 1 eq)in acetonitrile (40 mL) was added CH₂12 (8.98 g, 33.54 mmol, 2.71 mL,1.1 eq) and isopentyl nitrite (17.86 g, 152.45 mmol, 20.53 mL, 5.0 eq).The mixture was stirred at 80° C. for 3.5 h. LCMS showed the startingmaterial was consumed completely and one main peak was detected. Thereaction mixture was concentrated under reduced pressure to removeacetonitrile. The residue was diluted with EtOAc (50 mL), washed withNa₂SO₃ solution (50 mL×2), dried over Na₂SO₄, filtered and concentratedunder reduced pressure to give a residue which was purified by flashsilica gel chromatography (from PE/EtOAc=100/1 to 10/1, TLC:PE/EtOAc=3/1, R_(f)=0.90) to yield 4,6-dichloro-2-iodo-pyrimidine (6.32g, 22.60 mmol, 74.1% yield, 98.3% purity) as a white solid. ¹H NMR (400MHz, CD₃OD) δ ppm 7.79-7.76 (m, 1H); ES-LCMS m/z 274.7, 276.8 [M+H]⁺.

Step 2: 4,6-Dichloro-2-(5-fluoro-3-pyridyl)pyrimidine

4,6-Dichloro-2-iodo-pyrimidine (500 mg, 1.79 mmol, 1 eq),(5-fluoro-3-pyridyl)boronic acid (251.96 mg, 1.79 mmol, 1 eq), Na₂CO₃(568.55 mg, 5.36 mmol, 3.0 eq) and Pd(dppf)Cl₂ (130.84 mg, 178.81 umol,0.1 eq) in 1,4-dioxane (6 mL) and water (1.2 mL) were taken up into amicrowave tube. The sealed tube was heated at 80° C. for 30 min undermicrowave. LCMS showed 53% of desired compound was detected. Thereaction mixture was diluted with EtOAc (30 mL) and filtered through apad of celite. The filtrate was concentrated under reduced pressure togive a residue which was purified by flash silica gel chromatography(from PE/EtOAc=100/1 to 10/3, TLC: PE/EtOAc=3/1, R_(f)=0.80) to yield4,6-dichloro-2-(5-fluoro-3-pyridyl)pyrimidine (165 mg, 676.08 umol,37.8% yield, 100% purity) as a white solid. ¹H NMR (400 MHz, CD₃OD) δppm 9.36 (s, 1H), 8.67 (d, J=2.9 Hz, 1H), 8.52-8.44 (m, 1H), 7.76 (s,1H); ES-LCMS m/z 243.9, 245.9 [M+H]⁺.

Step 3:6-Chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine

To a solution of 4,6-dichloro-2-(5-fluoro-3-pyridyl)pyrimidine (165 mg,676.08 umol, 1.0 eq) in i-PrOH (10 mL) was added DIEA (262.13 mg, 2.03mmol, 353.28 uL, 3.0 eq) and 2-(1H-indol-3-yl)ethanamine (108.32 mg,676.08 umol, 1.0 eq). The mixture was stirred at 50° C. for 3 h. LCMSshowed 88% of desired compound was detected. The reaction mixture wasconcentrated to yield6-chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine(240 mg, crude) as brown oil which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.31-9.23 (m, 1H),8.86-8.20 (m, 2H), 8.11 (s, 1H), 7.92 (s, 1H), 7.57 (dd, J=7.8, 11.8 Hz,1H), 7.42-7.29 (m, 1H), 7.26-7.17 (m, 1H), 7.13-6.93 (m, 2H), 6.55 (s,1H), 3.83-3.70 (m, 2H), 3.04-2.95 (m, 2H); ES-LCMS m/z 368.0, 369.0[M+H]⁺.

Step 4:2-(5-Fluoro-3-pyridyl)-N6-[2-(1H-indol-3-yl)ethyl]-N4-isopropyl-pyrimidine-4,6-diamine(I-20)

6-Chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine(60 mg, 140.62 umol, 1.0 eq), propan-2-amine (1.03 g, 17.46 mmol, 1.5mL, 124.16 eq) and DIEA (90.87 mg, 703.10 umol, 122.46 uL, 5.0 eq) weretaken up into a microwave tube in i-PrOH (3 mL). The sealed tube washeated at 125° C. for 6 h under microwave. LCMS showed 66% of desiredcompound was detected. The reaction mixture was concentrated underreduced pressure to remove i-PrOH. The residue was purified bypreparative HPLC (HCl condition; column: Phenomenex Gemini 150×25 mm×10um; mobile phase: [water (0.05% HCl)-ACN]; B %: 25%-55%, 10 min) and thedesired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N6-[2-(1H-indol-3-yl)ethyl]-N4-isopropyl-pyrimidine-4,6-diamine(19.11 mg, 38.19 umol, 27.16% yield, 99.90% purity, 3HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.99 (s, 1H), 8.75 (d, J=2.6Hz, 1H), 8.18 (s, 1H), 7.62 (d, J=7.9 Hz, 1H), 7.31 (s, 1H), 7.17-6.98(m, 4H), 3.73 (s, 2H), 3.13 (t, J=6.4 Hz, 3H), 1.21 (s, 6H); ES-LCMS m/z391.3 [M+H]⁺.

Example 12

Synthesis of I-15

Synthetic Scheme:

Step 1: 1-Imidazo[1,2-a]pyridin-3-yl-N,N-dimethyl-methanamine

A mixture of imidazo[1,2-a]pyridine (500 mg, 4.23 mmol, 1 eq),N-methylmethanamine (414.16 mg, 5.08 mmol, 1.2 eq, HCl), and HCHO(412.16 mg, 5.08 mmol, 378.13 uL, 37% in water, 1.2 eq) in n-BuOH (5 mL)was degassed and purged with N₂ for 3 times. Then the mixture wasstirred at 120° C. for 3 h under N₂ atmosphere. LC-MS showed thestarting material was consumed completely and one main peak with desiredMS was detected. The reaction mixture was concentrated under reducedpressure to give 1-imidazo[1,2-a]pyridin-3-yl-N,N-dimethyl-methanamine(742 mg, crude) as a yellow solid, which was used in the next stepwithout further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.85 (d,J=6.8 Hz, 1H), 8.27 (d, J=6.8 Hz, 1H), 7.85-7.73 (m, 2H), 7.68 (s, 1H),4.41 (s, 2H), 2.64 (s, 6H); ES-LCMS m/z 176.2 [M+H]⁺.

Step 2: 2-Imidazo[1,2-a]pyridin-3-ylacetonitrile

To a solution of 1-imidazo[1,2-a]pyridin-3-yl-N,N-dimethyl-methanamine(742.00 mg, 4.23 mmol, 1 eq) in THE (6 mL) was added dimethyl sulfate(534.10 mg, 4.23 mmol, 401.58 uL, 1 eq) dropwise. The resulting mixturewas heated at 70° C. for 30 minutes. The solvent was removed and theresidue was dissolved in H₂O (5 mL). To the mixture was added KCN(303.30 mg, 4.66 mmol, 1.1 eq) and the mixture was heated at 110° C. for3 h. LC-MS showed 27% of the starting mixture was remained and 33% ofdesired compound was detected. The reaction mixture was quenched byaddition of NaHCO₃ (30 mL) then extracted with EtOAc (20 mL×3). Thecombined organic layers were washed with brine (10 mL), dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure togive a residue. The residue was purified by flash silica gelchromatography (from DCM/MeOH=1/0 to 10/1, TLC: DCM/MeOH=10/1,R_(f)=0.48) to yield 2-imidazo[1,2-a]pyridin-3-ylacetonitrile (900 mg,50.4% yield, 87.6% purity) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δppm 8.39 (d, J=6.8 Hz, 1H), 7.63 (d, J=9.0 Hz, 1H), 7.58 (s, 1H), 7.32(ddd, J=1.1, 6.7, 9.0 Hz, 1H), 7.06 (dt, J=0.9, 6.8 Hz, 1H), 4.47 (s,2H); ES-LCMS m/z 158.1 [M+H]⁺.

Step 3: 2-Imidazo[1,2-a]pyridin-3-ylethanamine

To a solution of 2-imidazo[1,2-a]pyridin-3-ylacetonitrile (80 mg, 421.70umol, 1 eq) in THE (8 mL) was added BH₃-Me₂S (10 M, 421.70 uL, 10 eq)dropwise. The mixture was stirred at 70° C. for 2 h. LC-MS showed thestarting material was consumed completely and desired MS was detected.The reaction mixture was quenched by addition of MeOH (20 mL) andHCl/MeOH (4 M, 0.1 mL) slowly followed by stirring at 70° C. for 3 h.The mixture was concentrated under reduced pressure to give2-imidazo[1,2-a]pyridin-3-ylethanamine (68 mg, crude) as colorless oil,which was used for the next step without further purification. ¹H NMR(400 MHz, CD₃OD) δ ppm 8.48-8.43 (m, 1H), 7.89-7.65 (m, 1H), 7.69-7.44(m, 1H), 7.46 (s, 1H), 7.26-7.17 (m, 1H), 3.17-3.10 (m, 2H), 3.09-3.01(m, 2H); ES-LCMS m/z 162.1 [M+H]⁺.

Step 4: 6-Chloro-9-isopropyl-purin-2-amine

To a solution of 6-chloro-9H-purin-2-amine (27 g, 159.22 mmol, 1 eq) inDMSO (220 mL) was added K₂CO₃ (66.02 g, 477.67 mmol, 3 eq) and2-bromopropane (97.92 g, 796.12 mmol, 74.75 mL, 5 eq). The mixture wasstirred at 15° C. for 88 h. LC-MS showed the starting material wasconsumed completely and one main peak with desired MS was detected. Thereaction mixture was quenched by addition of water (1000 mL) thenextracted with EtOAc (500 mL×3). The combined organic layers were washedwith water (100 mL×2), brine (100 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue as a yellow solid.The residue was added with PE/EtOAc (5/1, 500 mL) then stirred at 15° C.for 2 h. The slurry was filtered and the cake was rinsed with PE (30mL×2). The solid was collected and dried in vacuo to yield crude6-chloro-9-isopropyl-purin-2-amine (28 g, 130.97 mmol, 82.2% yield, 99%purity) as a light yellow solid, which was used for the next stepwithout further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.84 (s,1H), 5.20 (s, 2H), 4.74-4.64 (m, 1H), 1.57 (d, J=6.8 Hz, 6H); ES-LCMSm/z 212.0, 214.0 [M+H]⁺.

Step 5: 6-Chloro-2-iodo-9-isopropyl-purine

To a mixture of 6-chloro-9-isopropyl-purin-2-amine (10 g, 46.78 mmol, 1eq), I₂ (11.87 g, 46.78 mmol, 1 eq), CuI (8.91 g, 46.78 mmol, 1 eq), andCH₂I₂ (125.28 g, 467.75 mmol, 37.73 mL, 10 eq) in THE (400 mL) was addedisopentyl nitrite (16.44 g, 140.33 mmol, 18.89 mL, 3 eq). The mixturewas stirred at 70° C. for 3 h. LC-MS showed the starting material wasconsumed completely and desired MS was detected. The reaction mixturewas filtered through celite and the cake was rinsed with EtOAc (100mL×2). The filtrate was concentrated under reduced pressure and theresidue was diluted with EtOAc (300 mL), washed with Na₂SO₃ (100 mL),brine (100 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by flash silicagel chromatography (from PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=1/1,R_(f)=0.58) to yield 6-chloro-2-iodo-9-isopropyl-purine (12.5 g, 31.39mmol, 67.1% yield, 81% purity) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 8.09 (s, 1H), 4.98-4.82 (m, 1H), 1.63 (d, J=7.0 Hz, 6H);ES-LCMS m/z 322.8, 324.8 [M+H]⁺.

Step 6: 6-Chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine

6-Chloro-2-iodo-9-isopropyl-purine (3 g, 8.28 mmol, 1 eq),(5-fluoro-3-pyridyl)boronic acid (1.17 g, 8.28 mmol, 1 eq), Pd(dppf)Cl₂(302.86 mg, 413.91 umol, 0.05 eq) and Cs₂CO₃ (2.70 g, 8.28 mmol, 1 eq)in 1,4-dioxane (50 mL) and H₂O (10 mL) was de-gassed and then heated at80° C. for 16 h under N₂. LCMS showed the starting material was consumedcompletely. The reaction mixture was poured into H₂O (100 mL) thenextracted with EtOAc (80 mL×3). The organic phase was washed with brine(30 mL), dried over anhydrous Na₂SO₄, concentrated under reducedpressure to give a residue which was purified by flash silica gelchromatography (from PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=1/1, R_(f)=0.58)to yield 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine (1.71 g,5.63 mmol, 68.0% yield, 96% purity) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.54 (s, 1H), 8.58 (d, J=2.8 Hz, 1H), 8.48 (td, J=2.1, 9.5Hz, 1H), 8.21 (s, 1H), 5.10-4.93 (m, 1H), 1.73 (d, J=6.8 Hz, 6H);ES-LCMS m/z 292.0, 294.0 [M+H]⁺.

Step 7:2-(5-Fluoro-3-pyridyl)-N-(2-imidazo[1,2-a]pyridin-3-ylethyl)-9-isopropyl-purin-6-amine(I-15)

To a solution of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine (50mg, 164.55 umol, 1 eq), and 2-imidazo[1,2-a]pyridin-3-ylethanamine (68mg, 421.83 umol, 2.56 eq) in i-PrOH (5 mL) was added DIEA (106.33 mg,822.73 umol, 143.31 uL, 5 eq). The mixture was stirred at 95° C. for 16h. LC-MS showed the starting material was consumed completely anddesired MS (m/z=M/2+H) was detected. The reaction mixture wasconcentrated under reduced pressure and the residue was purified bypreparative HPLC (column: Phenomenex Kinetex XB-C18 150 mm×30 mm, 5 m;mobile phase: [water (0.05% HCl)-ACN]; B %: 7%-37%, 12 min) and thedesired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N-(2-imidazo[1,2-a]pyridin-3-ylethyl)-9-isopropyl-purin-6-amine(34.09 mg, 63.53 umol, 38.6% yield, 98% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.50 (s, 1H), 9.33 (s, 1H),9.03-8.88 (m, 3H), 8.00 (s, 1H), 7.96-7.88 (m, 1H), 7.86-7.79 (m, 1H),7.54 (t, J=6.8 Hz, 1H), 5.21-5.07 (m, 1H), 4.32 (t, J=6.2 Hz, 2H), 3.58(t, J=6.3 Hz, 2H), 1.74 (d, J=6.6 Hz, 6H); ES-LCMS m/z 416.9 [M+H]⁺.

Example 13

Synthesis of I-1

Synthetic Scheme:

Step 1: 3-Isopropyl-4-nitro-1H-pyrazole-5-carboxylic acid

To an ice-bath and stirred solution of fuming HNO₃ (700.00 mg, 11.11mmol, 0.5 mL, 1.71 eq) and fuming H₂SO₄ (1.88 g, 18.76 mmol, 1.02 mL,98% purity, 2.89 eq) was added 3-isopropyl-1H-pyrazole-5-carboxylic acid(1 g, 6.49 mmol, 1 eq) in portionwise at 0° C. over 5 min. Afteraddition, the mixture was stirred at this temperature for 1 h then at100° C. for 6 h. LC-MS showed the starting material was consumedcompletely and one main peak with desired MS was detected. The mixturewas poured into ice-water (30 g), the white precipitate was filtered anddried to yield 3-isopropyl-4-nitro-1H-pyrazole-5-carboxylic acid (700mg, 3.20 mmol, 49.36% yield, 91.1% purity) as a white solid. ¹H NMR (400MHz, CD₃OD) (5 ppm 3.56 (spt, J=7.0 Hz, 1H), 1.35 (d, J=7.1 Hz, 6H);ES-LCMS m/z 200.1 [M+H]⁺.

Step 2: 3-Isopropyl-4-nitro-1H-pyrazole-5-carboxamide

To a solution of 3-isopropyl-4-nitro-1H-pyrazole-5-carboxylic acid (700mg, 3.20 mmol, 1 eq) in DCM (15 mL) and DMF (0.1 mL) was added oxalylchloride (1.22 g, 9.61 mmol, 840.83 uL, 3 eq) dropwise over 5 min. Afteraddition, the mixture was stirred at 15° C. for 1 h then the resultingmixture was concentrated. The residue was dissolved in THE (10 mL) andcooled to 0° C. NH₃.H₂O (9.75 g, 77.90 mmol, 10.71 mL, 28% purity, 24.33eq) was added in dropwise. After addition, the mixture was stirred at15° C. for 1 h. LC-MS showed the starting material was consumedcompletely and one main peak with desired MS was detected. The reactionmixture was concentrated under reduced pressure and the residue wasdiluted with EtOAc (50 mL) and extracted with EtOAc (50 mL×2). Thecombined organic layers were washed with brine (30 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to give3-isopropyl-4-nitro-1H-pyrazole-5-carboxamide (0.6 g, 2.91 mmol, 90.96%yield, 96.2% purity) as a white solid. ¹H NMR (400 MHz, CD₃OD) δ ppm3.68-3.60 (m, 1H), 1.34 (d, J=6.8 Hz, 6H); ES-LCMS m/z 199.1 [M+H]⁺.

Step 3: 4-Amino-3-isopropyl-1H-pyrazole-5-carboxamide

To a mixture of 3-isopropyl-4-nitro-1H-pyrazole-5-carboxamide (600 mg,2.91 mmol, 1 eq) in MeOH (20 mL) was added Pd/C (10%, 0.1 g) under N₂.The suspension was degassed under vacuum and purged with H₂ severaltimes. The mixture was stirred under H₂ (15 psi) at 15° C. for 3 h. TLC(PE/EA=1/1, R_(f)=0.1) showed the starting material was consumedcompletely. The reaction mixture was filtered and the filtrate wasconcentrated to yield 4-amino-3-isopropyl-1H-pyrazole-5-carboxamide (500mg, 2.68 mmol, 91.86% yield, 90% purity) as an off white solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 3.02 (spt, J=7.0 Hz, 1H), 1.27 (d, J=7.1 Hz, 6H);ES-LCMS m/z 169.1 [M+H]⁺.

Step 4: 3-Isopropyl-1,4-dihydropyrazolo[4,3-d]pyrimidine-5,7-dione

To a solution of 4-amino-3-isopropyl-1H-pyrazole-5-carboxamide (500 mg,2.68 mmol, 1 eq) in DMF (10 mL) was added CDI (477.20 mg, 2.94 mmol, 1.1eq) and the mixture was stirred at 80° C. for 12 h. LC-MS showedstarting material was consumed completely and one main peak with desiredMS was detected. The reaction mixture was concentrated to yield3-isopropyl-1,4-dihydropyrazolo[4,3-d]pyrimidine-5,7-dione (550 mg,crude) as a brown solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 3.22-3.13 (m,1H), 1.30 (d, J=6.8 Hz, 6H); ES-LCMS m/z 195.1 [M+H]⁺.

Step 5: 5,7-Dichloro-3-isopropyl-1H-pyrazolo[4,3-d]pyrimidine

A solution of 3-isopropyl-1,4-dihydropyrazolo[4,3-d]pyrimidine-5,7-dione(500 mg, 2.57 mmol, 1 eq) in POCl₃ (10 mL) was stirred at 100° C. for 2h. LC-MS showed starting material was consumed completely. The reactionmixture was concentrated under reduced pressure. The residue was dilutedwith EtOAc (50 mL), adjusted pH to 8 and extracted with EtOAc (50 mL×2).The combined organic layers were washed with brine (30 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (ISCO®; 4g SepaFlash® Silica Flash Column, Eluent of 0˜30% Ethylacetate/Petroleum ether gradient @ 15 mL/min) to yield5,7-dichloro-3-isopropyl-1H-pyrazolo[4,3-d]pyrimidine (110 mg, 442.22umol, 17.18% yield, 92.9% purity) as a white solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 10.39 (br s, 1H), 3.51 (spt, J=6.9 Hz, 1H), 1.47 (d, J=6.8Hz, 6H); ES-LCMS m/z 231.3, 233.3 [M+H]⁺.

Step 6:5-Chloro-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-1H-pyrazolo[4,3-d]pyrimidin-7-amine

To a solution of 5,7-dichloro-3-isopropyl-1H-pyrazolo[4,3-d]pyrimidine(110 mg, 442.22 umol, 1 eq), 2-(1H-indol-3-yl)ethanamine (85.02 mg,530.67 umol, 1.2 eq) in i-PrOH (5 mL) was added DIEA (171.46 mg, 1.33mmol, 231.08 uL, 3 eq). The mixture was stirred at 8° C. for 16 h. LC-MSshowed starting material was consumed completely and one main peak withdesired MS was detected. The reaction mixture was concentrated underreduced pressure to give5-chloro-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-1H-pyrazolo[4,3-d]pyrimidin-7-amine(160 mg, 392.29 umol, 88.71% yield, 87% purity) as a brown solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 7.67 (br s, 1H), 7.35 (d, J=8.0 Hz, 1H),7.14-7.08 (m, 2H), 7.04-6.98 (m, 1H), 3.94 (q, J=6.2 Hz, 2H), 3.43 (brs, 1H), 3.17 (t, J=7.2 Hz, 2H), 1.39 (s, 6H); ES-LCMS m/z 355.2, 357.1[M+H]⁺.

Step 7:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-1H-pyrazolo[4,3-d]pyrimidin-7-amine(I-1)

To a mixture of5-chloro-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-1H-pyrazolo[4,3-d]pyrimidin-7-amine(80 mg, 196.15 umol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (33.17 mg,235.38 umol, 1.2 eq), Cs₂CO₃ (159.77 mg, 490.37 umol, 2.5 eq) in1,4-dioxane (2 mL) and water (0.5 mL) was added Pd(dppf)Cl₂ (7.18 mg,9.81 umol, 0.05 eq) under N₂. The mixture was stirred under N₂ at 120°C. for 30 min under microwave. LC-MS showed 82.5% of product wasdetected. The reaction mixture was concentrated under reduced pressureand the residue was diluted with DCM (20 mL) and extracted with DCM (20mL×2). The combined organic layers were washed with brine (30 mL), driedover Na₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: PhenomenexGemini 150*25 mm*10 um; mobile phase: [water (0.05% HCl)-ACN]; B %:40%-70%, 10 min). The desired fraction was lyophilized to yield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-1H-pyrazolo[4,3-d]pyrimidin-7-amine(66.84 mg, 123.15 umol, 62.78% yield, 96.7% purity, 3HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.93 (s, 1H), 8.74 (d, J=2.6Hz, 1H), 8.01 (d, J=8.8 Hz, 1H), 7.55 (d, J=7.9 Hz, 1H), 7.16 (d, J=7.9Hz, 1H), 7.04 (s, 1H), 6.94 (t, J=7.7 Hz, 1H), 6.88-6.81 (m, 1H), 4.18(t, J=6.6 Hz, 2H), 3.57 (d, J=7.0, 13.9 Hz, 1H), 3.21 (t, J=6.6 Hz, 2H),1.44 (d, J=7.1 Hz, 6H); ES-LCMS m/z 415.9 [M+H]⁺.

Example 14

Synthesis of I-9

Synthetic Scheme:

Step 1: N,N-Dimethyl-1-(1H-pyrrolo[2,3-c]pyridin-3-yl)methanamine

To a solution of 1H-pyrrolo[2,3-c]pyridine (900 mg, 7.62 mmol, 1 eq) inn-BuOH (15 mL) was added HCHO (680.06 mg, 8.38 mmol, 623.91 uL, 37% inwater, 1.1 eq) and N-methylmethanamine hydrochloride (695.78 mg, 8.53mmol, 1.12 eq). The mixture was stirred at 120° C. for 3 h. TLC(DCM/MeOH=10/1, R_(f)=0.06) indicated the starting material was consumedcompletely and one major new spot was detected. The reaction mixture wasconcentrated under reduced pressure to yieldN,N-dimethyl-1-(1H-pyrrolo[2,3-c]pyridin-3-yl)methanamine (1.85 g,crude) as a yellow oil which was used into the next step without furtherpurification. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.83 (d, J=1.1 Hz, 1H), 8.22(d, J=5.7 Hz, 1H), 7.97 (s, 1H), 7.91 (d, J=0.7 Hz, 1H), 4.56 (s, 2H),2.87 (s, 6H); ES-LCMS m/z 176.2 [M+H]⁺.

Step 2: 2-(1H-Pyrrolo[2,3-c]pyridin-3-yl)acetonitrile

To a solution ofN,N-dimethyl-1-(1H-pyrrolo[2,3-c]pyridin-3-yl)methanamine (2.75 g, 8.55mmol, 1 eq) in THE (20 mL) was added dimethyl sulfate (1.08 g, 8.55mmol, 810.81 uL, 1.0 eq) dropwise. The resulting mixture was stirred at70° C. for 30 min. THE was removed and the residue was dissolved inwater (15 mL). To the mixture was added KCN (612.44 mg, 9.40 mmol, 1.1eq) and the mixture was stirred at 110° C. for 3 h. LCMS showed thestarting material was consumed completely and one main peak with desiredMS was detected. The reaction mixture was diluted with water (40 mL) andextracted with EtOAc (45 mL×3). The combined organic layers were driedover Na₂SO₄, filtered and concentrated under reduced pressure to yield2-(1H-pyrrolo[2,3-c]pyridin-3-yl)acetonitrile (417.0 mg, crude) as ayellow solid which was used in the next step without furtherpurification. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.70 (s, 1H), 8.13 (d, J=5.8Hz, 1H), 7.67 (d, J=5.8 Hz, 1H), 7.58 (s, 1H), 4.01 (s, 2H); ES-LCMS m/z158.1 [M+H]⁺.

Step 3: 2-(1H-Pyrrolo[2,3-c]pyridin-3-yl)ethanamine

To a solution of 2-(1H-pyrrolo[2,3-c]pyridin-3-yl)acetonitrile (417.0mg, 2.65 mmol, 1 eq) in THF (20 mL) was added BH₃-Me₂S (10 M, 2.65 mL,10 eq). The mixture was stirred at 70° C. for 2 h. LCMS showed thestarting material was consumed completely and one main peak wasdetected. MeOH (50 mL) was added into the reaction mixture dropwise andit was stirred at 70° C. for 2 h. The mixture was concentrated underreduced pressure to yield 2-(1H-pyrrolo[2,3-c]pyridin-3-yl)ethanamine(423.9 mg, crude) as a yellow solid which was used into the next stepwithout further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.71-8.59(m, 1H), 8.08-8.00 (m, 1H), 7.73-7.64 (m, 1H), 7.63-7.53 (m, 1H),3.09-2.92 (m, 4H); ES-LCMS m/z 162.1 [M+H]⁺.

Step 4:2-(5-Fluoro-3-pyridyl)-9-isopropyl-N-[2-(1H-pyrrolo[2,3-c]pyridin-3-yl)ethyl]purin-6-amine(I-9)

To a solution of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine(49.35 mg, 148.88 umol, 1.0 eq) in i-PrOH (5 mL) was added DIEA (57.72mg, 446.64 umol, 77.79 uL, 3.0 eq) and2-(1H-pyrrolo[2,3-c]pyridin-3-yl)ethanamine (72.00 mg, 446.64 umol, 3.0eq). The mixture was stirred at 50° C. for 15 h. LCMS showed 71% ofdesired compound was detected. The reaction mixture was concentratedunder reduced pressure to remove i-PrOH. The residue was purified bypreparative HPLC (HCl condition; column: Phenomenex Gemini 150×25 mm×10um; mobile phase: [water (0.05% HCl)-ACN]; B %: 30%-60%, 10 min) and thedesired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-9-isopropyl-N-[2-(1H-pyrrolo[2,3-c]pyridin-3-yl)ethyl]purin-6-amine(11.84 mg, 21.76 umol, 14.62% yield, 96.66% purity, 3HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.54 (s, 1H), 9.36 (s, 1H),9.05-8.98 (m, 2H), 8.96 (s, 1H), 8.27-8.22 (m, 1H), 8.21-8.16 (m, 2H),5.15 (td, J=6.7, 13.6 Hz, 1H), 4.21 (t, J=6.4 Hz, 2H), 3.39 (t, J=6.7Hz, 2H), 1.74 (d, J=6.8 Hz, 6H); ES-LCMS m/z 417.0 [M+H]⁺.

Example 15

Synthesis of I-11

Synthetic Scheme:

Step 1:N-(2-(5,7-Difluoro-2-methyl-1H-indol-3-yl)ethyl)-2-(5-fluoropyridin-3-yl)-9-isopropyl-9H-purin-6-amine(I-11)

To a solution of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine (50mg, 150.83 umol, 1 eq) in i-PrOH (2 mL) was added DIEA (97.47 mg, 754.17umol, 131.36 uL, 5 eq) and2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethanamine (45.29 mg, 150.83umol, 1 eq, oxalic acid). The mixture was stirred at 50° C. for 16 h.LCMS showed 65% of desired compound was detected. The reaction mixturewas concentrated under reduced pressure to give a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150×25 mm×10 um;mobile phase: [water (0.05% HCl)-ACN]; B %. 50%-80%, 10 min) and thedesired fraction was lyophilized to yieldN-(2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethyl)-2-(5-fluoropyridin-3-yl)-9-isopropyl-9H-purin-6-amine(53.81 mg, 36.8% yield, 100% purity, 3HCl salt) as a light yellow solid.¹H NMR (400 MHz, CD₃OD) δ ppm 9.36 (s, 1H), 8.85 (s, 1H), 8.76 (s, 1H),8.61 (d, J=8.8 Hz, 1H), 6.95 (d, J=7.3 Hz, 1H), 6.49 (t, J=10.2 Hz, 1H),5.11-4.99 (m, 1H), 4.03 (s, 2H), 3.08 (t, J=6.5 Hz, 2H), 2.31 (s, 3H),1.70 (d, J=6.8 Hz, 6H); ES-LCMS m/z 466.0 [M+H]⁺.

Example 16

Synthesis of I-14

Synthetic Scheme:

Step 1: tert-Butyl (2-(1H-indol-3-yl)ethyl)carbamate

To a solution of 2-(1H-indol-3-yl)ethanamine (2.8 g, 17.48 mmol, 1 eq)in anhydrous DCM (30 mL) was added TEA (5.31 g, 52.44 mmol, 7.30 mL, 3eq) and (Boc)₂O (4.58 g, 20.98 mmol, 4.82 mL, 1.2 eq). The mixture wasstirred at 15° C. for 3 h. LCMS showed the starting material wasconsumed completely and one main peak with desired MS was detected. H₂O(20 mL) was added, and the mixture was extracted with DCM (20 mL×3). Thecombined organic layers were washed with brine (20 mL×2), dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure togive a residue which was purified on silica gel column chromatography(from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=2/1, R_(f)=0.45) to yieldtert-butyl (2-(1H-indol-3-yl)ethyl)carbamate (3.55 g, 13.50 mmol, 77.2%yield, 99% purity) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.13(br s, 1H), 7.61 (d, J=7.9 Hz, 1H), 7.35 (d, J=8.2 Hz, 1H), 7.24-7.16(m, 1H), 7.16-7.08 (m, 1H), 7.00 (s, 1H), 4.59 (br s, 1H), 3.47 (q,J=6.4 Hz, 2H), 2.96 (t, J=6.8 Hz, 2H), 1.46 (s, 9H); ES-LCMS m/z 283.0[M+Na]⁺.

Step 2: tert-Butyl (2-(1-methyl-1H-indol-3-yl)ethyl)carbamate

To a solution of tert-butyl N-[2-(1H-indol-3-yl)ethyl]carbamate (1.5 g,5.70 mmol, 1 eq) in THE (30 mL) was added NaH (456.30 mg, 11.41 mmol,60% purity, 2.0 eq) at 0° C. over 5 min. After addition, the mixture wasstirred at this temperature for 20 min then MeI (809.67 mg, 5.70 mmol,355.12 uL, 1.0 eq) was added dropwise at 0° C. The resulting mixture wasstirred at 10° C. for 2 h. TLC (PE/EtOAc=3/1, R_(f)=0.49) indicated thestarting material was consumed completely and two new spots formed. Thereaction mixture was quenched by addition of water (50 mL) at 10° C.then extracted with EtOAc (30 mL×3). The combined organic layers werewashed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedby flash silica gel chromatography (from PE/EA=1/0 to 3/1, TLC:PE/EA=3/1, R_(f)=0.49) to yield tert-butylN-[2-(1-methylindol-3-yl)ethyl]carbamate (900 mg, 50.4% yield, 87.6%purity) as a colorless oil; ¹H NMR (400 MHz, CD₃OD) δ ppm 7.98 (d, J=7.8Hz, 1H), 7.74-7.57 (m, 2H), 7.55-7.43 (m, 1H), 4.99 (br s, 1H), 4.13 (s,3H), 3.89-3.81 (m, 2H), 3.32 (t, J=6.4 Hz, 2H), 1.82 (s, 9H); ES-LCMSm/z 219.0 [M-t-Bu+H]⁺.

Step 3: 2-(1-Methyl-1H-indol-3-yl)ethanamine

To a solution of tert-butyl N-[2-(1-methylindol-3-yl)ethyl]carbamate(700 mg, 2.24 mmol, 1 eq) in DCM (20 mL) was added HCl/MeOH (4 M, 5 mL,8.95 eq). The mixture was stirred at 10° C. for 4 h. TLC (EtOAc,R_(f)=0.0) indicated the start material was consumed completely. Thereaction mixture was concentrated under reduced pressure to give crude2-(1-methylindol-3-yl)ethanamine (500 mg, crude, 2HCl) as a white solidwhich was used for the next step without further purification. ¹H NMR(400 MHz, CD₃OD) δ ppm 7.58 (d, J=7.8 Hz, 1H), 7.36 (d, J=8.3 Hz, 1H),7.20 (t, J=7.5 Hz, 1H), 7.14-7.04 (m, 2H), 3.78 (s, 3H), 3.27-3.19 (m,2H), 3.15-3.07 (m, 2H); ES-LCMS m/z 175.1 [M+H]⁺.

Step 4:2-Chloro-9-isopropyl-N-(2-(1-methyl-1H-indol-3-yl)ethyl)-9H-purin-6-amine

To a solution of 2,6-dichloro-9-isopropyl-purine (200 mg, 827.24 umol, 1eq) in i-PrOH (10 mL) was added DIEA (855.30 mg, 6.62 mmol, 1.15 mL, 8eq) and 2-(1-methylindol-3-yl)ethanamine (306.70 mg, 992.69 umol, 1.2eq, 2HCl). The mixture was stirred at 50° C. for 14 h. LC-MS showed thestarting material was consumed completely and one main peak with desiredMS was detected. The reaction mixture was concentrated under reducedpressure to remove i-PrOH (10 mL) and the residue was purified by flashsilica gel chromatography (from PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=1/2,R_(f)=0.62) to yield2-chloro-9-isopropyl-N-[2-(1-methylindol-3-yl)ethyl]purin-6-amine (250mg, 81.9% yield, 100% purity) as colorless oil. ¹H NMR (400 MHz, CD₃OD)δ ppm 8.48-8.19 (m, 2H), 7.73 (d, J=7.5 Hz, 1H), 7.37 (d, J=8.3 Hz, 1H),7.25-6.94 (m, 3H), 4.70-4.58 (m, 1H), 3.77-3.64 (m, 5H), 3.06-2.91 (m,2H), 1.50 (d, J=6.5 Hz, 6H); ES-LCMS m/z 369.1, 371.1 [M+H]⁺.

Step 5:2-(5-Fluoropyridin-3-yl)-9-isopropyl-N-(2-(1-methyl-1H-indol-3-yl)ethyl)-9H-purin-6-amine(I-14)

2-Chloro-9-isopropyl-N-[2-(1-methylindol-3-yl)ethyl]purin-6-amine (100mg, 271.10 umol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (95.50 mg,677.76 umol, 2.5 eq), Pd(dppf)Cl₂ (19.84 mg, 27.11 umol, 0.1 eq) andCs₂CO₃ (264.99 mg, 813.31 umol, 3.0 eq) were taken up into a microwavetube in 1,4-dioxane (3 mL) and H₂O (0.6 mL). The sealed tube was heatedat 120° C. for 30 min under microwave. LC-MS showed the start materialwas consumed completely and one main peak with desired MS was detected.The reaction mixture was concentrated under reduced pressure. Theresidue was dissolved in EtOAc (50 mL), filtered through SiO₂. The cakewas rinsed with EtOAc (10 mL×2). The combined organic layers wereconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um; mobilephase: [water (0.05% HCl)-ACN]; B %: 55%-85%, 10 min). The desiredfraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-9-isopropyl-N-[2-(1-methylindol-3-yl)ethyl]purin-6-amine(53.81 mg, 36.8% yield, 100% purity, 3HCl salt) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 9.36 (s, 1H), 9.11 (br s, 1H), 8.84 (s, 1H),8.75 (d, J=8.8 Hz, 1H), 7.60 (d, J=7.9 Hz, 1H), 7.20 (d, J=7.9 Hz, 1H),7.07 (t, J=7.5 Hz, 1H), 7.02-6.93 (m, 2H), 5.09 (td, J=6.8, 13.6 Hz,1H), 4.16-4.10 (m, 2H), 3.61 (s, 3H), 3.20 (t, J=6.6 Hz, 2H), 1.72 (d,J=6.8 Hz, 6H); ES-LCMS m/z 430.2 [M+H]⁺.

Example 17

Synthesis of I-7

Synthetic Scheme:

Step 1. N,N-Dimethyl-1-(1H-pyrrolo[3,2-b]pyridin-3-yl)methanamine

To a solution of 1H-pyrrolo[3,2-b]pyridine (500 mg, 4.23 mmol, 1 eq) inn-BuOH (5 mL) was added HCHO (377.65 mg, 4.65 mmol, 346.47 uL, 37% inwater, 1.1 eq) and N-methylmethanamine (379.43 mg, 4.65 mmol, 1.1 eq,HCl salt). The mixture was stirred at 120° C. for 3 h. LC-MS showedstarting material was consumed completely and one main peak with desiredMS was detected. The reaction mixture was concentrated under reducedpressure to giveN,N-dimethyl-1-(1H-pyrrolo[3,2-b]pyridin-3-yl)methanamine (559.7 mg,crude) as a yellow solid which was used in the next step without furtherpurification. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.32 (dd, J=1.1, 4.6 Hz,1H), 7.80 (d, J=8.3 Hz, 1H), 7.56 (s, 1H), 7.16 (dd, J=4.8, 8.3 Hz, 1H),3.80 (s, 2H), 2.28 (s, 6H); ES-LCMS m/z 176.1 [M+H]⁺.

Step 2: 2-(1H-Pyrrolo[3,2-b]pyridin-3-yl)acetonitrile

To a solution ofN,N-dimethyl-1-(1H-pyrrolo[3,2-b]pyridin-3-yl)methanamine (200.00 mg,1.14 mmol, 1 eq) in THE (1.5 mL) was added dimethyl sulfate (143.96 mg,1.14 mmol, 108.24 uL, 1.0 eq) dropwise. The resulting mixture wasstirred at 70° C. for 30 min. The solvent was removed and the residuewas dissolved in water (1.2 mL). To the mixture was added KCN (81.76 mg,1.26 mmol, 1.1 eq) and the mixture was stirred at 110° C. for 3 h. LC-MSshowed starting material was consumed completely and one main peak withdesired MS was detected. The reaction mixture was diluted with water (5mL) and extracted with EtOAc (15 mL×3). The combined organic layers weredried over Na₂SO₄, filtered and concentrated under reduced pressure togive 2-(1H-pyrrolo[3,2-b]pyridin-3-yl)acetonitrile (131.7 mg, 662.72umol, 58.1% yield, 79.1% purity) as a yellow solid which was used in thenext step without further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm8.37-8.29 (m, 1H), 7.87-7.78 (m, 1H), 7.64-7.51 (m, 1H), 7.25-7.14 (m,1H), 4.01 (d, J=0.9 Hz, 2H); ES-LCMS m/z 158.1 [M+H]⁺.

Step 3: 2-(1H-Pyrrolo[3,2-b]pyridin-3-yl)ethanamine

To a solution of 2-(1H-pyrrolo[3,2-b]pyridin-3-yl)acetonitrile (131.70mg, 662.72 umol, 1 eq) in THF (10 mL) was added BH₃-Me₂S (1 mL, 10 M inMe₂S) dropwise. The reaction mixture was stirred at 70° C. for 2 h.LC-MS showed starting material was consumed completely and one main peakwas detected. MeOH (30 mL) was added dropwise and the mixture wasstirred at 70° C. for 2 h. The mixture was concentrated under reducedpressure to give 2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethanamine (50.00 mg,crude) as a yellow solid which was used in the next step without furtherpurification. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.30 (d, J=4.6 Hz, 1H), 7.80(d, J=8.2 Hz, 1H), 7.44 (s, 1H), 7.16 (dd, J=4.6, 8.2 Hz, 1H), 3.35 (s,2H), 3.03 (s, 2H); ES-LCMS m/z 162.2 [M+H]⁺.

Step 4:2-Chloro-9-isopropyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]purin-6-amine

To a solution of 2,6-dichloro-9-isopropyl-purine (76.49 mg, 320.46 umol,1.2 eq) in i-PrOH (10 mL) was added DIEA (172.57 mg, 1.34 mmol, 232.57uL, 5.0 eq) and 2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethanamine (50.00 mg,267.05 umol, 1 eq). The mixture was stirred at 50° C. for 12 h. LC-MSshowed starting material was consumed completely and 55% of desiredcompound was detected. The reaction mixture was concentrated and theresidue was purified by preparative TLC (SiO₂, DCM/MeOH=10/1,R_(f)=0.65) to yield2-chloro-9-isopropyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]purin-6-amine(25.00 mg, 70.26 umol, 26.3% yield, 100% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 8.30 (dd, J=1.3, 4.8 Hz, 1H), 8.08 (s, 1H),7.77 (dd, J=1.1, 8.2 Hz, 1H), 7.44 (s, 1H), 7.14 (dd, J=4.8, 8.3 Hz,1H), 4.79-4.68 (m, 1H), 3.96-3.87 (m, 2H), 3.20 (t, J=6.8 Hz, 2H), 1.56(d, J=6.8 Hz, 6H); ES-LCMS m/z 356.1, 358.1 [M+H]⁺.

Step 5:2-(5-Fluoro-3-pyridyl)-9-isopropyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]purin-6-amine(I-7)

2-Chloro-9-isopropyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]purin-6-amine(25.00 mg, 70.26 umol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (24.75mg, 175.65 umol, 2.5 eq), Cs₂CO₃ (68.68 mg, 210.78 umol, 3.0 eq) andPd(dppf)Cl₂ (5.14 mg, 7.03 umol, 0.1 eq) were taken up into a microwavetube in 1,4-dioxane (2 mL) and water (0.4 mL). The sealed tube washeated at 120° C. for 30 min under microwave. LC-MS showed startingmaterial was consumed completely and 60% of desired compound wasdetected. The reaction mixture was diluted with EtOAc (10 mL) andfiltered through a pad of celite. The filtrate was concentrated underreduced pressure to give a residue which was purified by preparativeHPLC (HCl condition, column: Phenomenex Kinetex XB-C18 150 mm*30 mm, 5m; mobile phase: [water (0.05% HCl)-ACN]; B %: 5%-35%, 12 min) and thedesired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-9-isopropyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]purin-6-amine(8.83 mg, 16.14 umol, 23.0% yield, 96.1% purity, 3HCl salt) as a whitesolid; ¹H NMR (400 MHz, CD₃OD) δ ppm 9.37 (s, 1H), 8.92 (s, 1H), 8.78(s, 1H), 8.69 (s, 1H), 8.51 (s, 1H), 8.43 (d, J=7.3 Hz, 1H), 8.05 (s,1H), 7.59 (s, 1H), 5.06 (td, J=6.8, 13.6 Hz, 1H), 4.22 (t, J=6.4 Hz,2H), 3.38 (t, J=6.8 Hz, 2H), 1.70 (d, J=6.8 Hz, 6H); ES-LCMS m/z 417.2[M+H]⁺.

Example 18

Synthesis of I-13

Synthetic Scheme:

Step 1:N-(2-(1H-Benzo[d]imidazol-1-yl)ethyl)-2-chloro-9-isopropyl-9H-purin-6-amine

To a solution of 2,6-dichloro-9-isopropyl-purine (200 mg, 827.24 umol, 1eq) in i-PrOH (8 mL) was added DIEA (106.91 mg, 827.24 umol, 144.09 uL,1 eq) and 2-(benzimidazol-1-yl)ethanamine (160.02 mg, 992.69 umol, 1.2eq). The mixture was stirred at 50° C. for 16 h. LC-MS showed thestarting material was consumed completely and one main peak with desiredMS was detected. The reaction mixture was concentrated under reducedpressure to remove i-PrOH to give a residue which was purified by flashsilica gel column chromatography (from DCM/MeOH=1/0 to 10/1, TLC:DCM/MeOH=10/1, R_(f)=0.33) to yieldN-[2-(benzimidazol-1-yl)ethyl]-2-chloro-9-isopropyl-purin-6-amine (275mg, 93.4% yield, 100% purity) as a white solid. ¹H NMR (400 MHz, CD₃OD)δ ppm 7.98-7.71 (m, 2H), 7.54 (d, J=7.5 Hz, 2H), 7.38-7.18 (m, 2H), 6.77(br s, 1H), 4.86-4.67 (m, 1H), 4.53 (t, J=6.0 Hz, 2H), 4.02 (br s, 2H),1.69-1.46 (m, 6H); ES-LCMS m/z 356.1, 357.1 [M+H]⁺.

Step 2:N-(2-(1H-Benzo[d]imidazol-1-yl)ethyl)-2-(5-fluoropyridin-3-yl)-9-isopropyl-9H-purin-6-amine(I-13)

N-[2-(Benzimidazol-1-yl)ethyl]-2-chloro-9-isopropyl-purin-6-amine (120mg, 337.24 umol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (95.04 mg,674.49 umol, 2 eq), Cs₂CO₃ (329.64 mg, 1.01 mmol, 3.0 eq) andPd(dppf)Cl₂ (12.34 mg, 16.86 umol, 0.05 eq) in 1,4-dioxane (5 mL) andH₂O (1 mL) was de-gassed and refilled with N₂. The mixture was heated to80° C. for 12 h. LC-MS showed 58% of desired compound. The reactionmixture was poured into H₂O (30 mL) then extracted with EtOAc (20 mL×3).The organic phase was combined, washed with brine (10 mL), dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to give aresidue. The residue was purified by preparative HPLC (column:Phenomenex Gemini 150*25 mm*10 um; mobile phase: [water (0.05% ammoniahydroxide v/v)-ACN]; B %: 38%-68%, 10 min) to yieldN-[2-(benzimidazol-1-yl)ethyl]-2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-amine(31.40 mg, 22.4% yield, 100% purity) as a white solid. ¹H NMR (400 MHz,CD₃OD) δ ppm 9.08 (br s, 1H), 8.43 (d, J=2.6 Hz, 1H), 8.17 (s, 1H),8.15-7.95 (m, 2H), 7.67 (d, J=8.2 Hz, 1H), 7.38 (d, J=8.2 Hz, 1H), 7.26(t, J=7.7 Hz, 1H), 7.17-7.00 (m, 1H), 4.88-4.83 (m, 1H), 4.62 (t, J=5.4Hz, 2H), 4.20 (br s, 2H), 1.63 (d, J=6.8 Hz, 6H); ES-LCMS m/z 417.1[M+H]⁺.

Example 19

Synthesis of I-8

Synthetic Scheme:

Step 1: 2,6-Dichloro-9-isopropyl-purine

To a solution of 2,6-dichloro-9H-purine (20 g, 105.82 mmol, 1 eq) inDMSO (160 mL) was added K₂CO₃ (73.13 g, 529.09 mmol, 5 eq) and2-bromopropane (65.07 g, 529.09 mmol, 49.67 mL, 5 eq). The mixture wasstirred at 25° C. for 12 h. TLC (PE/EtOAc=1/1, R_(f)=0.45) indicatedstarting material was consumed completely and one major new spot wasdetected. The reaction mixture was diluted with water (200 mL) andextracted with EtOAc (100 mL×3). The combined organic layers were washedwith water (80 mL×3), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by flash silicagel column chromatography (from PE/EtOAc=5/1 to 2/1, TLC: PE/EtOAc=1/1,R_(f)=0.45) to yield 2,6-dichloro-9-isopropyl-purine (3.52 g, 14.75mmol, 13.9% yield, 96.8% purity) as a white solid. ¹H NMR (400 MHz,CD₃OD) δ ppm 8.66 (s, 1H), 4.97-4.89 (m, 1H), 1.65 (d, J=6.8 Hz, 6H);ES-LCMS m/z 231.0, 232.9 [M+H]⁺.

Step 2:2-Chloro-9-isopropyl-N-[2-(1H-pyrrolo[3,2-c]pyridin-3-yl)ethyl]purin-6-amine

To a solution of 2,6-dichloro-9-isopropyl-purine (148.07 mg, 620.33umol, 1 eq) in i-PrOH (10 mL) was added DIEA (400.86 mg, 3.10 mmol,540.24 uL, 5.0 eq) and 2-(1H-pyrrolo[3,2-c]pyridin-3-yl)ethanamine (100mg, 620.33 umol, 1 eq). The mixture was stirred at 50° C. for 12 h.LC-MS showed starting material was consumed completely and 33% ofdesired compound was detected. The reaction mixture was concentratedunder reduced pressure to remove solvent. The residue was purified byflash silica gel chromatography (from DCM/MeOH=100/1 to 5/1, TLC:DCM/MeOH=10/1, R_(f)=0.68) to yield2-chloro-9-isopropyl-N-[2-(1H-pyrrolo[3,2-c]pyridin-3-yl)ethyl]purin-6-amine(74.5 mg, 33.8% yield, crude) as a yellow solid; ES-LCMS m/z 356.0,357.0 [M+H]⁺.

Step 3:2-(5-Fluoro-3-pyridyl)-9-isopropyl-N-[2-(1H-pyrrolo[3,2-c]pyridin-3-yl)ethyl]purin-6-amine(I-8)

2-Chloro-9-isopropyl-N-[2-(1H-pyrrolo[3,2-c]pyridin-3-yl)ethyl]purin-6-amine(74.5 mg, 104.69 umol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (36.88mg, 261.72 umol, 2.5 eq), Cs₂CO₃ (102.33 mg, 314.06 umol, 3.0 eq) andPd(dppf)Cl₂ (7.66 mg, 10.47 umol, 0.1 eq) were taken up into a microwavetube in 1,4-dioxane (3 mL) and water (0.6 mL). The sealed tube washeated at 120° C. for 30 min under microwave. LC-MS showed startingmaterial was consumed completely, 56% of desired compound was detected.The reaction mixture was diluted with EtOAc (15 mL) and filtered througha pad of celite. The filtrate was concentrated under reduced pressure togive a residue which was purified by preparative HPLC (HCl condition,column: Phenomenex Gemini 150×25 mm×10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 15%-45%, 10 min) and the desired fraction waslyophilized to yield2-(5-fluoro-3-pyridyl)-9-isopropyl-N-[2-(1H-pyrrolo[3,2-c]pyridin-3-yl)ethyl]purin-6-amine(22.19 mg, 42.20 umol, 40.3% yield, 100% purity, 3HCl salt) as a whitesolid; ¹H NMR (400 MHz, CD₃OD) δ ppm 9.53 (s, 1H), 9.31 (s, 2H),9.00-8.94 (m, 2H), 8.28 (d, J=6.5 Hz, 1H), 7.85 (d, J=6.8 Hz, 1H), 7.79(s, 1H), 5.14 (d, J=6.7, 13.5 Hz, 1H), 4.23 (s, 2H), 3.41 (t, J=6.7 Hz,2H), 1.74 (d, J=6.8 Hz, 6H); ES-LCMS m/z 417.2 [M+H]⁺.

Example 20

Synthesis of I-21

Synthetic Scheme:

Step 1: 4,6-Dichloro-2-(5-fluoro-3-pyridyl)pyrimidine

4,6-Dichloro-2-iodo-pyrimidine (1 g, 3.58 mmol, 1 eq),(5-fluoro-3-pyridyl)boronic acid (503.91 mg, 3.58 mmol, 1 eq), Na₂CO₃(1.14 g, 10.73 mmol, 3.0 eq) and Pd(dppf)Cl₂ (261.67 mg, 357.62 umol,0.1 eq) were taken up into a microwave tube in 1,4-dioxane (14 mL) andwater (2.4 mL). The sealed tube was heated at 80° C. for 30 min undermicrowave. LCMS showed 58% of desired compound was detected. Thereaction mixture was diluted with EtOAc (50 mL) and filtered through apad of celite. The filtrate was concentrated under reduced pressure togive a residue which was purified by flash silica gel chromatography(from PE/EtOAc=100/1 to 5/1, TLC: PE/EtOAc=3/1, R_(f)=0.80) to yield4,6-dichloro-2-(5-fluoro-3-pyridyl)pyrimidine (275 mg, 1.13 mmol, 37.8%yield, 100% purity) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm9.34 (t, J=1.4 Hz, 1H), 8.66 (d, J=2.9 Hz, 1H), 8.51-8.43 (m, 1H), 7.75(s, 1H); ES-LCMS m/z 243.9, 245.9 [M+H]⁺.

Step 2: 4-Chloro-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidine

To a solution of i-PrOH (14.78 mg, 245.85 umol, 18.82 uL, 1 eq) in THE(4 mL) was added NaH (11.80 mg, 295.02 umol, 60% in mineral oil, 1.2eq). The mixture was stirred at 0° C. for 30 min.4,6-dichloro-2-(5-fluoro-3-pyridyl)pyrimidine (60 mg, 245.85 umol, 1.0eq) was added into the above solution and the mixture was stirred at 15°C. for 12 h. LCMS showed 83% of desired compound was detected. Thereaction mixture was concentrated under reduced pressure to give aresidue which was diluted with water (30 mL) and extracted with EtOAc(30 mL×3). The combined organic layers were dried over Na₂SO₄, filteredand concentrated under reduced pressure to yield4-chloro-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidine (50 mg, 167.92umol, 68.3% yield, 89.9% purity) as a brown solid, which was used in thenext step without further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm9.39-9.28 (m, 1H), 8.68-8.58 (m, 1H), 8.50-8.39 (m, 1H), 6.85 (s, 1H),5.57 (m, 1H), 1.44 (d, J=6.2 Hz, 6H); ES-LCMS m/z 268.0, 270.0 [M+H]⁺.

Step 3:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-4-amine(I-21)

To a solution of 4-chloro-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidine(50 mg, 167.92 umol, 1.0 eq) in i-PrOH (4 mL) was added DIEA (65.11 mg,503.76 umol, 87.74 uL, 3.0 eq) and 2-(1H-indol-3-yl)ethanamine (134.52mg, 839.60 umol, 5 eq). The mixture was stirred at 70° C. for 12 h. LCMSshowed the starting materials were remained and there was no desiredcompound. The reaction mixture was added into a microwave tube andheated at 125° C. for 3 h under microwave. LCMS showed the startingmaterial was consumed completely and 28% of desired compound wasdetected. The reaction mixture was concentrated under reduced pressureto give a residue which was purified by preparative HPLC (HCl condition;column: Phenomenex Gemini 150×25 mm×10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 55%-85%, 10 min) and the desired fraction waslyophilized to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-4-amine(26.35 mg, 52.45 umol, 31.2% yield, 99.7% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.15 (s, 1H), 8.66 (d, J=2.4 Hz,1H), 8.32 (d, J=8.8 Hz, 1H), 7.60 (d, J=7.7 Hz, 1H), 7.29 (d, J=7.9 Hz,1H), 7.13-6.95 (m, 3H), 5.69 (m, 1H), 5.04 (m, 1H), 3.78 (m, 2H), 3.09(t, J=6.8 Hz, 2H), 1.40-1.33 (m, 6H); ES-LCMS m/z 392.1 [M+H]⁺.

Example 21

Synthesis of I-23

Synthetic Scheme:

Step 1: tert-Butyl N-(2-bromo-4,6-difluoro-phenyl)carbamate

A mixture of 2-bromo-4,6-difluoro-aniline (5 g, 24.04 mmol, 1 eq), Boc₂O(15.74 g, 72.11 mmol, 16.57 mL, 3 eq), DMAP (293.67 mg, 2.40 mmol, 0.1eq) in THE (50 mL) was degassed and purged with N₂ for 3 times, and thenthe mixture was stirred at 70° C. for 16 h under N₂ atmosphere. LC-MSshowed the starting material was consumed completely and the di-BOCintermediate was detected. The reaction mixture was concentrated underreduced pressure to give a residue which was dissolved in MeOH (50 mL)and K₂CO₃ (9.97 g, 72.11 mmol, 3 eq) was added. The mixture was stirredat 70° C. for 4 h. LC-MS showed the intermediate was consumed completelyand one main peak with desired MS was detected. The reaction mixture wasfiltered, the filtrate was concentrated under reduced pressure. To theresidue was added water (100 mL), extracted with EtOAc (60 mL×3). Thecombined organic layers were washed with brine (30 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue. To the residue was added n-heptane (100 mL) then was stirredfor 1 h at 15° C. The slurry was filtered, the cake was rinsed withn-heptane (30 mL×2), dried in vacuo to yield tert-butylN-(2-bromo-4,6-difluoro-phenyl)carbamate (4.15 g, 13.47 mmol, 56.0%yield, 100% purity) as a white solid, which was used in the next stepwithout further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.18 (td,J=2.3, 7.7 Hz, 1H), 6.89 (dt, J=2.8, 8.9 Hz, 1H), 5.91 (s, 1H), 1.51 (s,9H); ES-LCMS m/z 251.9, 253.9 [M-t-Bu+H]⁺.

Step 2: tert-ButylN-[2,4-difluoro-6-(2-trimethylsilylethynyl)phenyl]carbamate

A mixture of tert-butyl N-(2-bromo-4,6-difluoro-phenyl)carbamate (4 g,12.98 mmol, 1 eq), ethynyl(trimethyl)silane (2.55 g, 25.96 mmol, 3.60mL, 2.0 eq), TEA (3.94 g, 38.95 mmol, 5.42 mL, 3.0 eq), CuI (247.24 mg,1.30 mmol, 0.1 eq) and Pd(PPh₃)₂Cl₂ (455.60 mg, 649.10 umol, 0.05 eq) inDMF (80 mL) was degassed and purged with N₂ for 3 times, and then themixture was stirred at 100° C. for 16 h under N₂ atmosphere. LC-MSshowed the starting material was consumed completely and desired MS wasdetected. The reaction mixture was quenched by addition water (300 mL),extracted with EtOAc (200 mL×3). The combined organic layers were washedwith brine (50 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedby flash silica gel chromatography (from PE/EtOAc=1/0 to 10/1, TLC:PE/EtOAc=20/1, R_(f)=0.31) to yield tert-butylN-[2,4-difluoro-6-(2-trimethylsilylethynyl)phenyl]carbamate (2.05 g,5.04 mmol, 38.8% yield, 80% purity) as a black brown solid. ¹H NMR (400MHz, CDCl₃) δ ppm 7.00-6.94 (m, 1H), 6.90-6.81 (m, 1H), 6.10 (s, 1H),1.51 (s, 9H), 0.27 (m, 9H); ES-LCMS m/z 270.0 [M-t-Bu+H]⁺.

Step 3: 5,7-Difluoro-1H-indole

To EtOH (100 mL) was added Na (791.23 mg, 34.42 mmol, 8 eq) slowly.After being stirred for 1 h at 15° C., tert-butylN-[2,4-difluoro-6-(2-trimethylsilylethynyl)phenyl]carbamate (1.75 g,4.30 mmol, 1 eq) was added to the above solution. The mixture wasstirred at 85° C. for 16 h. LC-MS showed the starting material wasconsumed completely and desired MS was detected. The reaction mixturewas concentrated under reduced pressure to remove EtOH. To the residuewas added water (100 mL) and extracted with EtOAc (60 mL×3). Thecombined organic layers were washed with brine (30 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 10/1, TLC: PE/EtOAc=20/1, R_(f)=0.17) to yield5,7-difluoro-1H-indole (410 mg, 1.87 mmol, 43.5% yield, 70% purity) asblack oil. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.31 (s, 1H), 7.29-7.22 (m,1H), 7.09 (dd, J=2.1, 9.2 Hz, 1H), 6.73 (ddd, J=2.2, 9.4, 11.1 Hz, 1H),6.59-6.51 (m, 1H); ES-LCMS: No correct mass was found.

Step 4: 5,7-Difluoro-1H-indole-3-carbaldehyde

To a solution of DMF (10 mL) was added POCl₃ (350.46 mg, 2.29 mmol,212.40 uL, 2.0 eq) dropwise at −20° C. over a period of 10 mins underN₂. After being stirred for 1 h, 5,7-difluoro-1H-indole (250 mg, 1.14mmol, 1 eq) in DMF (2 mL) was added to the above solution during whichthe temperature was maintained below −20° C. The reaction mixture waswarmed to 15° C. and stirred for 1 h. LC-MS showed the starting materialwas consumed completely and desired MS was detected. The reactionmixture was quenched by addition NaHCO₃ (30 mL), extracted with EtOAc(20 mL×3). The combined organic layers were washed with brine (10 mL),dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to give a residue which was purified by flash silica gelchromatography (from PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=1/1, R_(f)=0.40)to yield 5,7-difluoro-1H-indole-3-carbaldehyde (200 mg, 993.71 umol,86.9% yield, 90% purity) as a yellow solid. ¹H NMR (400 MHz, Acetone) δppm 11.74 (s, 1H), 10.06 (d, J=0.8 Hz, 1H), 8.38 (s, 1H), 7.74 (dd,J=2.3, 9.0 Hz, 1H), 7.03 (ddd, J=2.3, 9.5, 11.3 Hz, 1H); ES-LCMS m/z182.1 [M+H]⁺.

Step 5: 5,7-Difluoro-3-[(E)-2-nitrovinyl]-1H-indole

To a solution of 5,7-difluoro-1H-indole-3-carbaldehyde (200 mg, 993.71umol, 1 eq) in nitronethane (8 mL) was added NH₄O Ac (229.79 mg, 2.98mmol, 3.0 eq). The mixture was stirred at 110° C. for 16 h. LC-MS showedthe starting material was consumed completely and desired MS wasdetected. The reaction mixture was concentrated under reduced pressureto remove nitronethane. The residue was diluted with EtOAc (50 mL),washed with water (10 mL), brine (10 mL), dried over Na₂SO₄, filteredand concentrated under reduced pressure to give5,7-difluoro-3-[(E)-2-nitrovinyl]-1H-indole (160 mg, 599.56 umol, 60.3%yield, 84% purity) as a brown solid, which was used in the next stepwithout further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.23 (d,J=13.6 Hz, 1H), 7.74-7.58 (m, 2H), 7.26-7.11 (m, 1H), 6.91-6.75 (m, 1H);ES-LCMS m/z 225.0 [M+H]⁺.

Step 6: 2-(5,7-Difluoro-1H-indol-3-yl)ethanamine

To a solution of 5,7-difluoro-3-[(E)-2-nitrovinyl]-1H-indole (50 mg,187.36 umol, 1 eq) in THF (5 mL) was added dropwise LAH (1 M, 936.82 uL,5 eq) at 0° C. After addition, the mixture was stirred at 80° C. for 2h. LC-MS showed the starting material was consumed completely and onemain peak with desired MS was detected. The reaction mixture was dilutedwith THE (50 mL), quenched by addition water (0.05 mL), follow by 10%NaOH (0.05 mL) and water (0.15 mL) in sequence at 0° C. After beingstirred for 30 min, the mixture was filtered through celite. Thefiltrate was concentrated under reduced pressure to give2-(5,7-difluoro-1H-indol-3-yl)ethanamine (36 mg, crude) as a yellow oil,which was used in the next step without further purification. ¹H NMR(400 MHz, CD₃OD) δ ppm 7.18 (s, 1H), 7.13-7.01 (m, 1H), 6.73-6.67 (m,1H), 2.97-2.79 (m, 4H); ES-LCMS m/z 197.2 [M+H]⁺.

Step 7:N-[2-(5,7-Difluoro-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-amine(I-23)

To a solution of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine (50mg, 164.55 umol, 1 eq) in i-PrOH (3 mL) was added DIEA (106.33 mg,822.73 umol, 143.30 uL, 5 eq) and2-(5,7-difluoro-1H-indol-3-yl)ethanamine (35.51 mg, 181.00 umol, 1.1eq). The mixture was stirred at 60° C. for 16 h. LC-MS showed 21% of thestarting material was remained and 66% of desired compound was detected.The reaction mixture was concentrated under reduced pressure to removei-PrOH to give a residue which was purified by preparative HPLC (column:Phenomenex Gemini 150×25 mm×10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 46%-76%, 10 min). The desired fraction was lyophilizedto yieldN-[2-(5,7-difluoro-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-amine(20.82 mg, 36.75 umol, 22.3% yield, 99% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.45 (s, 1H), 9.20 (s, 1H), 8.90(s, 1H), 8.83 (d, J=9.0 Hz, 1H), 7.24 (s, 1H), 7.11 (d, J=8.8 Hz, 1H),6.67-6.58 (m, 1H), 5.11 (m, 1H), 4.13 (s, 2H), 3.17 (t, J=6.6 Hz, 2H),1.73 (d, J=6.8 Hz, 6H); ES-LCMS m/z 452.2 [M+H]⁺.

Example 22

Synthesis of I-16

Synthetic Scheme:

Step 1: 5-Fluoropyridine-3-carbonyl chloride

To a solution of 5-fluoropyridine-3-carboxylic acid (7 g, 49.61 mmol, 1eq) in SOCl₂ (57.40 g, 482.47 mmol, 35.00 mL, 9.73 eq) was added DMF(0.1 mL) at 0° C. After addition, the mixture was stirred at 50° C. for2 h. TLC (PE/EA=1/1, R_(f)=0.75, added MeOH) showed the startingmaterial was consumed completely. The mixture was concentrated to yieldthe 5-fluoropyridine-3-carbonyl chloride (7 g, crude) as light yellowoil, which was used in the next step without further purification.

Step 2: 5-Fluoropyridine-3-carboxamide

To a solution of NH₃.H₂O (31.85 g, 254.47 mmol, 35.00 mL, 28% purity,5.80 eq) in THE (10 mL) was added the solution of5-fluoropyridine-3-carbonyl chloride (7 g, 43.87 mmol, 1 eq) in THE (30mL) dropwise at 0° C. under N₂. The mixture was stirred at 15° C. for 1h. LCMS showed starting material was consumed completely and one mainpeak with desired MS was detected. The reaction mixture was concentratedunder reduced pressure to remove solvent. The residue was diluted withEtOAc (50 mL) and extracted with EtOAc (50 mL×2). The combined organiclayers were washed with brine (30 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to yield5-fluoropyridine-3-carboxamide (6 g, 42.39 mmol, 96.6% yield, 99.0%purity) as an off white solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.87 (s,1H), 8.62 (d, J=2.6 Hz, 1H), 8.06-8.01 (m, 1H); ES-LCMS m/z 141.1[M+H]⁺.

Step 3: 5-Fluoropyridine-3-carbonitrile

To a mixture of 5-fluoropyridine-3-carboxamide (6 g, 42.39 mmol, 1 eq)and TEA (6.43 g, 63.59 mmol, 8.85 mL, 1.5 eq) in DCM (60 mL) was addedTFAA (13.36 g, 63.59 mmol, 8.85 mL, 1.5 eq) dropwise at 15° C. under N₂.The mixture was stirred at 15° C. for 12 h. TLC (PE/EA=1/1, R_(f)=0.89)indicated the starting material was consumed completely and one new spotformed. The reaction mixture was concentrated under reduced pressure togive a residue which was purified by flash silica gel chromatography(ISCO®; 24 g SepaFlash Silica Flash Column, Eluent of 0˜20% Ethylacetate/Petroleum ethergradient @ 30 mL/min) to yield5-fluoropyridine-3-carbonitrile (4.7 g, 36.57 mmol, 86.3% yield, 95.0%purity) as a white solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.82-8.71 (m,2H), 8.14-8.06 (m, 1H); ES-LCMS: No correct mass was found.

Step 4: 5-Fluoro-N-(4-isopropyl-1H-pyrazol-5-yl)pyridine-3-carboxamidine

A mixture of 5-fluoropyridine-3-carbonitrile (1.02 g, 7.91 mmol, 1 eq)and 4-isopropyl-1H-pyrazol-5-amine (880.27 mg, 6.33 mmol, 0.8 eq) inxylene (20 mL) was stirred for 30 min at 70° C. Then AlMe₃ (2 M, 4.75mL, 1.2 eq) was added to above mixture in one portion at 70° C. underN₂. The mixture was stirred at 100° C. for 15 h. LC-MS showed 16% ofdesired MS was detected. The mixture was quenched by MeOH (20 mL),concentrated to yield a residue which was purified by flash silica gelchromatography (ISCO®; 12 g SepaFlash Silica Flash Column, Eluent of0˜10% MeOH/DCM ethergradient @ 30 mL/min) to yield5-fluoro-N-(4-isopropyl-1H-pyrazol-5-yl)pyridine-3-carboxamidine (600mg, 1.94 mmol, 24.5% yield, 80% purity) as a yellow solid. ¹H NMR (400MHz, CD₃OD) δ ppm 8.96 (s, 1H), 8.54 (d, J=2.6 Hz, 1H), 8.11 (d, J=9.5Hz, 1H), 7.34 (s, 1H), 3.06 (m, 1H), 1.25 (d, J=6.8 Hz, 6H); ES-LCMS m/z248.1 [M+H]⁺.

Step 5:2-(5-Fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-ol

To a mixture of5-fluoro-N-(4-isopropyl-1H-pyrazol-5-yl)pyridine-3-carboxamidine (100mg, 323.53 umol, 1 eq) in 1,4-dioxane (10 mL) and THE (5 mL) was addedtriphosgene (33.60 mg, 113.24 umol, 0.35 eq) and then the mixture wasstirred for 15 h at 80° C. LC-MS showed 57.5% of desired MS wasdetected. The reaction mixture was concentrated under reduced pressureto give2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-ol(88 mg, crude) as a brown solid, which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.90-8.85 (m, 2H),8.18 (d, J=2.3, 8.6 Hz, 1H), 7.66 (s, 1H), 3.04 (d, J=6.9, 13.7 Hz, 1H),1.26 (d, J=6.8 Hz, 6H); ES-LCMS m/z 274.3 [M+H]⁺.

Step 6:4-Chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine

A mixture of2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-ol(88 mg, 322.03 umol, 1 eq) in POCl₃ (8.1 g, 52.83 mmol, 4.91 mL, 164.04eq) was stirred for 2 h at 100° C. LC-MS showed the starting materialwas consumed completely. The reaction mixture was concentrated underreduced pressure to give a residue, then diluted with DCM (20 mL) andextracted with DCM (20 mL×2). The combined organic layers were washedwith brine (15 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by flash silicagel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluentof 0˜15% Ethyl acetate/Petroleum ethergradient @ 30 mL/min) to yield4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(15 mg, 49.78 umol, 15.5% yield, 96.8% purity) as a white solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.49 (s, 1H), 8.60 (d, J=2.6 Hz, 1H), 8.42 (d,J=9.5 Hz, 1H), 8.17 (s, 1H), 3.35 (d, J=6.7, 13.9 Hz, 1H), 1.43 (d,J=6.8 Hz, 6H); ES-LCMS m/z 292.0, 294.0 [M+H]⁺.

Step 7:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-16)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(15 mg, 49.78 umol, 1 eq), DIEA (19.30 mg, 149.33 umol, 26.01 uL, 3 eq)and 2-(1H-indol-3-yl)ethanamine (9.57 mg, 59.73 umol, 1.2 eq) in i-PrOH(3 mL) was stirred for 3 h at 50° C. LC-MS showed the starting materialwas consumed completely. The reaction mixture was concentrated underreduced pressure to give a residue which was purified by preparativeHPLC (HCl condition; column: Phenomenex Kinetex XB-C18 150 mm*30 mm, 5μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 55%-85%, 17 min) and thedesired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-amine(6.17 mg, 11.70 umol, 23.5% yield, 99.5% purity, 3HCl) as a white solid.¹H NMR (400 MHz, CD₃OD) δ ppm 9.14 (s, 1H), 8.81 (br s, 1H), 8.56 (d,J=9.0 Hz, 1H), 7.97 (s, 1H), 7.69-7.63 (m, 1H), 7.18-7.12 (m, 1H),7.00-6.94 (m, 3H), 4.04 (t, J=6.7 Hz, 2H), 3.27-3.20 (m, 1H), 3.17 (t,J=6.7 Hz, 2H), 1.37 (d, J=6.8 Hz, 6H); ES-LCMS m/z 416.2 [M+H]⁺.

Example 23

Synthesis of I-24

Synthetic Scheme:

Step 1:[5-(5-Fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-3-yl]-morpholino-methanone(I-24)

To a solution of5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (25 mg, 55.60 μmol, 1 eq) in DCM (3 mL) was added HATU (31.71 mg,83.40 μmol, 1.5 eq), TEA (11.25 mg, 111.20 μmol, 15.48 uL, 2 eq) andmorpholine (9.69 mg, 111.20 μmol, 9.79 μL, 2 eq). The mixture wasstirred at 25° C. for 12 h under N₂ atmosphere. The mixture wasconcentrated under reduced pressure, then water (30 mL) was added,extracted with EtOAc (20 mL×3). The combined organic layers were washedwith brine (30 mL), dried over Na₂SO₄, filtered and concentrated. Theresidue was purified by preparative HPLC (column: Phenomenex Gemini150×25 mm×10 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 40%-70%, 10min) followed by lyophilization to yield[5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-3-yl]-morpholino-methanone(8.45 mg, 13.61 μmol, 24.4% yield, 100% purity, 3HCl) as a yellow solid.¹H NMR (400 MHz, CD₃OD) δ ppm 9.24 (s, 1H), 8.66 (d, J=2.6 Hz, 1H),8.55-8.48 (m, 1H), 8.34 (s, 1H), 7.39 (d, J=7.7 Hz, 1H), 7.28 (d, J=7.9Hz, 1H), 7.10-7.01 (m, 2H), 6.99-6.93 (m, 1H), 4.54 (br s, 1H), 3.80 (s,8H), 3.37-3.34 (m, 1H), 3.20-3.05 (m, 1H), 3.04-2.87 (m, 2H), 2.36 (brs, 1H), 2.28 (d, J=6.2 Hz, 1H); ES-LCMS m/z 512.3 [M+H]⁺.

Example 24

Synthesis of I-25

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-methoxy-N-methyl-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxamide(I-25)

To a solution of5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (30 mg, 63.06 μmol, 1 eq) in DCM (4 mL) was added HATU (35.97 mg,94.59 μmol, 1.5 eq), TEA (12.76 mg, 126.12 μmol, 17.55 μL, 2 eq) andN-methoxymethanamine (12.30 mg, 126.12 μmol, 2 eq, HCl). The mixture wasstirred at 20° C. for 1 h under N₂ atmosphere. The mixture wasconcentrated under reduced pressure, then water (30 mL) was added. Themixture was extracted with EtOAc (20 mL×3). The combined organic layerswere washed with brine (30 mL), dried over Na₂SO₄, filtered andconcentrated. The mixture was purified by preparative HPLC (column:Phenomenex Gemini 150×25 mm×10 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 40%-70%, 10 min) followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-N-methoxy-N-methyl-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxamide(15.98 mg, 26.86 μmol, 42.6% yield, 100% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.10 (s, 1H), 8.82 (d, J=2.8 Hz,1H), 8.67 (s, 1H), 8.47 (d, J=9.3 Hz, 1H), 7.39 (d, J=7.5 Hz, 1H),7.35-7.25 (m, 2H), 7.05 (t, J=7.0 Hz, 1H), 7.00-6.91 (m, 1H), 4.69 (brs, 1H), 3.87 (s, 3H), 3.43 (s, 3H), 3.18-2.95 (m, 4H), 2.37 (dd, J=5.6,10.2 Hz, 2H); ES-LCMS m/z 486.2 [M+H]⁺.

Example 25

Synthesis of I-26

Synthetic Scheme:

Step 1:(3R)—N-(3-isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidin-7-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-26)

A mixture of 7-chloro-3-isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidine (60mg, 218.15 μmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(44.69 mg, 239.96 μmol, 1.1 eq), DIEA (84.58 mg, 654.44 μmol, 113.99 μL,3 eq) in i-PrOH (2 mL) was degassed and purged with N₂ for 3 times, thenthe mixture was stirred at 80° C. for 12 h under N₂ atmosphere. Thereaction mixture was concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: PhenomenexGemini C18 250*50 mm*10 um; mobile phase: [water (0.05% HCl)-ACN]; B %:35%-65%, 10 min) followed by lyophilization to yield(3R)—N-(3-isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidin-7-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(45.09 mg, 91.19 μmol, 41.8% yield, 100.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.12 (s, 1H), 7.97 (d, J=7.7 Hz,2H), 7.68-7.50 (m, 3H), 7.39 (d, J=7.9 Hz, 1H), 7.27 (d, J=8.2 Hz, 1H),7.09-7.01 (m, 1H), 7.00-6.92 (m, 1H), 6.78 (s, 1H), 4.56 (m, 1H),3.39-3.33 (m, 2H), 3.16-2.90 (m, 3H), 2.53-2.07 (m, 2H), 1.39 (d, J=6.8Hz, 6H); ES-LCMS m/z 422.2 [M+H]⁺.

Example 26

Synthesis of I-27a

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[(5S)-4,5,6,7-tetrahydro-1H-benzimidazol-5-yl]pyrazolo[1,5-a]pyrimidin-7-amine(I-27)

A mixture of 4,5,6,7-tetrahydro-1H-benzimidazol-5-amine (62 mg, 451.95μmol, 1 eq),7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(145.99 mg, 451.95 μmol, 1 eq) and DIEA (175.24 mg, 1.36 mmol, 236.17μL, 3 eq) in i-PrOH (3 mL) was stirred at 90° C. for 12 h. The reactionmixture was concentrated under reduced pressure to dryness to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=20/0 to 0/1, TLC: PE/EtOAc=1/1, R_(f)=0.10). The desiredfraction was dried under reduced pressure to dryness. The residue wasseparated by preparative SFC (column: AD (250 mm*30 mm, Sum); mobilephase: [0.1% NH₃H₂O IPA]; B %: 35%-35%, min) to give Peak 1 (Rt=5.226)and Peak 2 (Rt=5.531). Peak 1 was concentrated under reduced pressure todryness to give a residue which was purified by preparative HPLC(column: Phenomenex Gemini 150*25 mm*10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 20%-50%, 10 min). The desired fraction was lyophilizedto give5-(5-fluoro-3-pyridyl)-3-isopropyl-N-[(5S)-4,5,6,7-tetrahydro-1H-benzimidazol-5-yl]pyrazolo[1,5-a]pyrimidin-7-amine(25.12 mg, 50.16 μmol, 11.1% yield, 100.0% purity, 3HCl) (Rt=5.226,[α]^(25.5) _(D)=−18.573 (0.105 g/100 mL in MeOH, ee %=97.4%) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.08 (s, 1H), 8.83 (s, 1H), 8.77(s, 1H), 8.47 (d, J=8.4 Hz, 1H), 8.25 (s, 1H), 7.07 (s, 1H), 4.72 (br s,1H), 3.45-3.31 (m, 2H), 3.07-2.87 (m, 3H), 2.44-2.35 (m, 1H), 2.33-2.22(m, 1H), 1.39 (d, J=6.8 Hz, 6H); ES-LCMS m/z 392.2 [M+H]⁺.

Example 27

Synthesis of I-28a

Synthetic Scheme:

Step 1: 2-(4-Methoxyphenyl)propanenitrile

To a stirred solution of 1-(4-methoxyphenyl)ethanol (2 g, 13.14 mmol, 1eq) in DCM (10 mL) was added the solution of TMSCN (2.61 g, 26.28 mmol,3.29 mL, 2 eq) and tribromoindigane (465.90 mg, 1.31 mmol, 0.1 eq) inDCM (10 mL) dropwise over 30 min. Then the resulted mixture was stirredat 30° C. for 15 min. TLC (PE/EtOAc=10/1, R_(f)=0.55) showed desiredcompound was detected. The reaction mixture was concentrated underreduced pressure to give a residue which was purified by flash silicagel chromatography (from PE/EtOAc=100/1 to 10/1, TLC: PE/EtOAc=10/1,R_(f)=0.55) to give 2-(4-methoxyphenyl)propanenitrile (1 g, 5.58 mmol,42.5% yield, 90.0% purity) as yellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm7.27 (s, 2H), 6.97-6.86 (m, 2H), 3.91-3.79 (m, 4H), 1.63 (d, J=7.3 Hz,3H).

Step 2: 2-(4-Methoxyphenyl)propan-1-amine

To a solution of 2-(4-methoxyphenyl)propanenitrile (800 mg, 4.47 mmol, 1eq) in MeOH (40 mL) was added Raney-Ni (0.5 g). The mixture was degassedand purged with H₂ three times and the mixture was stirred at 30° C. for5 h under H₂ atmosphere. TLC (PE/EtOAc=10/1, R_(f)=0.10) showed reactant1 was almost consumed and one new spot was detected. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by flash silica gel chromatography (from DCM/MeOH=10/1 to5/1, TLC: DCM/MeOH=10/1, R_(f)=0.60) to give2-(4-methoxyphenyl)propan-1-amine (710 mg, 3.87 mmol, 86.6% yield, 90.0%purity) as yellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.13 (d, J=8.5 Hz,2H), 6.87 (d, J=8.5 Hz, 2H), 3.80 (s, 3H), 2.88-2.69 (m, 3H), 1.23 (d,J=6.8 Hz, 3H); ES-LCMS m/z 166.1 [M+H]⁺.

Step 3:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[2-(4-methoxyphenyl)propyl]pyrazolo[1,5-a]pyrimidin-7-amine

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(250 mg, 773.93 μmol, 1.0 eq) in i-PrOH (10 mL) was added DIEA (300.07mg, 2.32 mmol, 404.41 μL, 3.0 eq) and 2-(4-methoxyphenyl)propan-1-amine(200 mg, 1.09 mmol, 1.41 eq). The mixture was stirred at 60° C. for 5 h.The reaction mixture was concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=100/1 to 3/1, TLC: PE/EtOAc=3/1, R_(f)=0.75) to give5-(5-fluoro-3-pyridyl)-3-isopropyl-N-[2-(4-methoxyphenyl)propyl]pyrazolo[1,5-a]pyrimidin-7-amine(300 mg, 715.15 μmol, 92.4% yield, 100.0% purity) as yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.02 (d, J=1.5 Hz, 1H), 8.53 (d, J=2.9 Hz,1H), 8.19-8.12 (m, 1H), 7.86 (s, 1H), 7.22 (d, J=8.6 Hz, 2H), 6.94-6.86(m, 2H), 6.38 (t, J=5.7 Hz, 1H), 6.19 (s, 1H), 3.79 (s, 3H), 3.68-3.53(m, 2H), 3.39-3.29 (m, 1H), 3.22-3.13 (m, 1H), 1.47-1.39 (m, 9H);ES-LCMS m/z 420.2 [M+H]⁺.

Step 4:4-[(1S)-2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-methyl-ethyl]phenol(I-28)

5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[2-(4-methoxyphenyl)propyl]pyrazolo[1,5-a]pyrimidin-7-amine(290.00 mg, 691.31 μmol, 1 eq) was added into HBr (25 mL, 60% in water).The mixture was stirred at 120° C. for 2 h. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby flash silica gel chromatography (from PE/EtOAc=100/1 to 2/1, TLC:PE/EtOAc=3/1, R_(f)=0.50). The compounds were separated by SFC(condition: column: OJ (250 mm×30 mm, 5 um); mobile phase: [0.1% NH3H2OETOH]; B %: 25%-25%, min). The solution after separation wereconcentrated to afford the crude products which were purified bypreparative HPLC (HCl condition; column: Phenomenex Gemini 150×25 mm×10μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 45%-75%, 10 min),followed by lyophilization to yield4-[(1S)-2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-methyl-ethyl]phenol(26.29 mg, 54.96 μmol, 7.9% yield, 100.0% purity, 2HCl salt (Rt=4.768min, ee %=100.0 and [α]²⁵ _(D)=+81.522 (MeOH, c=0.104 g/100 mL)) asyellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.81-8.79 (m, 2H), 8.24 (s,1H), 8.11 (td, J=2.2, 9.0 Hz, 1H), 7.12-7.06 (m, 2H), 6.62-6.56 (m, 2H),6.30 (s, 1H), 3.90-3.79 (m, 2H), 3.30-3.25 (m, 1H), 3.19-3.11 (m, 1H),1.41 (d, J=7.1 Hz, 3H), 1.36 (dd, J=2.5, 6.9 Hz, 6H); ES-LCMS m/z 406.2[M+H]⁺.

Example 28

Synthesis of I-29

Synthetic Scheme:

Step 1: Methyl5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of methyl7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(300 mg, 929.31 μmol, 1 eq) in i-PrOH (20 mL) was added DIEA (360.32 mg,2.79 mmol, 485.61 μL, 3 eq) and(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (190.39 mg, 1.02 mmol, 1.1eq). The mixture was stirred at 60° C. for 3 h under N₂ atmosphere. Themixture was concentrated under reduced pressure, then water (80 mL) wasadded. The mixture was extracted with EtOAc (50 mL×3). The combinedorganic layers were washed with brine (50 mL), dried over Na₂SO₄,filtered and concentrated. The residue was purified by flash silica gelchromatography (from PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=1/1, R_(f)=0.6)to yield a product of methyl5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxylate(270 mg, 561.92 μmol, 60.4% yield, 95% purity) as a yellow solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 9.09 (s, 1H), 8.58 (d, J=2.6 Hz, 1H), 8.47 (s,1H), 8.37-8.25 (m, 1H), 7.95 (s, 1H), 7.47 (d, J=7.9 Hz, 1H), 7.35 (d,J=7.9 Hz, 1H), 7.22-7.06 (m, 2H), 6.79 (d, J=8.6 Hz, 1H), 6.66 (s, 1H),4.36 (br s, 1H), 3.97 (s, 3H), 3.41 (dd, J=4.5, 15.3 Hz, 1H), 3.07-2.88(m, 3H), 2.42-2.27 (m, 2H); ES-LCMS m/z 457.2 [M+H]⁺.

Step 2: Methyl5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of methyl5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxylate(220 mg, 457.86 μmol, 1 eq) in H₂O (4 mL), MeOH (2 mL) and THE (2 mL)was added LiOH H₂O (275.60 mg, 6.57 mmol, 14.34 eq). The mixture wasstirred at 50° C. for 12 h under N₂ atmosphere. The residue wasdissolved in water (30 mL), adjusted to pH to 6 by 1N HCl, thenextracted with EtOAc (40 mL×3). The combined organic layers were washedwith brine (50 mL), dried over Na₂SO₄, filtered and concentrated. Theresidue was concentrated under reduced pressure to give methyl5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (150 mg, 315.29 μmol, 68.8% yield, 93.0% purity)) as a yellow solidwhich was used in the next step without further purification. ¹H NMR(400 MHz, CD₃OD) δ ppm 9.25 (s, 1H), 8.59-8.50 (m, 2H), 8.49-8.40 (m,1H), 7.38 (d, J=7.8 Hz, 1H), 7.26 (d, J=8.0 Hz, 1H), 7.09 (s, 1H),7.06-6.87 (m, 2H), 4.50 (s, 1H), 3.28 (s, 1H), 3.15-3.02 (m, 1H),3.00-2.83 (m, 2H), 2.43-2.31 (m, 1H), 2.30-2.15 (m, 1H); ES-LCMS m/z443.1 [M+H]⁺.

Step 3:[5-(5-Fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-3-yl]-(4-methylpiperazin-1-yl)methanone(I-29)

To a solution of5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (50 mg, 105.10 μmol, 1 eq) in DCM (5 mL) was added HATU (59.94 mg,157.65 μmol, 1.5 eq) and TEA (21.27 mg, 210.20 μmol, 29.26 μL, 2 eq) and1-methylpiperazine (15.79 mg, 157.65 μmol, 17.49 μL, 1.5 eq). Themixture was stirred at 20° C. for 1 h under N₂ atmosphere. The mixturewas concentrated under reduced pressure, then water (20 mL) was added.The mixture was extracted with EtOAc (20 mL×3). The combined organiclayers were washed with brine (30 mL), dried over Na₂SO₄, filtered andconcentrated. The residue was purified by preparative HPLC (column:Phenomenex Gemini C18 250×50 mm×10 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 10%-40%, 10 min) followed by lyophilization to yield[5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-3-yl]-(4-methylpiperazin-1-yl)methanone(24.27 mg, 36.20 μmol, 34.4% yield, 100% purity, 4HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.41 (s, 1H), 9.01-8.88 (m, 2H),8.43 (s, 1H), 7.37 (d, J=7.7 Hz, 1H), 7.27 (d, J=8.2 Hz, 1H), 7.22 (s,1H), 7.04 (t, J=7.5 Hz, 1H), 6.99-6.93 (m, 1H), 4.79-4.51 (m, 3H), 3.60(d, J=11.9 Hz, 4H), 3.29 (s, 3H), 3.20-3.07 (m, 1H), 3.03-2.90 (m, 5H),2.43-2.21 (m, 2H); ES-LCMS m/z 525.3 [M+H]⁺.

Example 29

Synthesis of I-30

Synthetic Scheme:

Step 1:(3R)—N-(6-Chloro-3-isopropyl-imidazo[1,2-a]pyrazin-8-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine

To a solution of 6,8-dichloro-3-isopropyl-imidazo[1,2-a]pyrazine (45 mg,195.57 μmol, 1 eq) and (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(38.25 mg, 205.35 μmol, 1.05 eq) in i-PrOH (5 mL) was added DIEA (75.83mg, 586.72 μmol, 102.20 μL, 3 eq). The mixture was stirred at 60° C. for3 h. LC-MS showed 55% of starting material was remained and 5% ofdesired compound was detected. The mixture was stirred at 60° C. for 16h. LC-MS showed 34% of starting material was remained and 30% of desiredcompound was detected. The mixture was stirred at 60° C. for 16 h. LC-MSshowed 21% of starting material was remained and 53% of desired compoundwas detected. The mixture was stirred at 60° C. for 16 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified on silica gel column chromatography (from PE/EtOAc=1/0 to10/3, TLC: PE/EtOAc=3/1, R_(f)=0.45) to give the product(3R)—N-(6-chloro-3-isopropyl-imidazo[1,2-a]pyrazin-8-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(60 mg, 157.94 μmol, 80.8% yield, 100.0% purity) was obtained as lightyellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.80 (s, 1H), 7.45 (d, J=7.5Hz, 1H), 7.32 (s, 1H), 7.22 (s, 1H), 7.17-7.12 (m, 1H), 7.12-7.06 (m,1H), 6.28 (d, J=9.3 Hz, 1H), 4.81 (br s, 1H), 3.29 (dd, J=5.4, 16.0 Hz,1H), 3.15-3.04 (m, 1H), 2.99-2.89 (m, 2H), 2.81 (dd, J=6.9, 15.1 Hz,1H), 2.28-2.25 (m, 1H), 2.23-2.15 (m, 1H), 1.37 (d, J=6.8 Hz, 6H);ES-LCMS m/z 380.2 [M+H]⁺.

Step 2:(3R)—N-[6-(5-Fluoro-3-pyridyl)-3-isopropyl-imidazo[1,2-a]pyrazin-8-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-30)

To a solution of(3R)—N-(6-chloro-3-isopropyl-imidazo[1,2-a]pyrazin-8-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(60 mg, 157.94 μmol, 1 eq) and (5-fluoro-3-pyridyl)boronic acid (44.51mg, 315.89 μmol, 2.0 eq) in 1,4-dioxane (2 mL) and H₂O (0.5 mL) wasadded Pd(dppf)Cl₂ (11.56 mg, 15.79 μmol, 0.1 eq) and Cs₂CO₃ (154.38 mg,473.83 μmol, 3.0 eq). The sealed tube was purged with N₂ for 3 min andheated at 110° C. for 0.5 h under microwave. The reaction mixture wasfiltered and concentrated under reduced pressure to give a residue whichwas purified by preparative HPLC twice (column: Phenomenex Gemini 150*25mm*10 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 40%-70%, 10 min;column: Gemini 150*25 5 u; mobile phase: [water (0.05% ammonia hydroxidev/v)-ACN]; B %: 50%-80%, 10 min), followed by lyophilization to yieldcompound(3R)—N-[6-(5-fluoro-3-pyridyl)-3-isopropyl-imidazo[1,2-a]pyrazin-8-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(26.85 mg, 60.59 μmol, 38.4% yield, 99.4% purity) as a white solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 9.07 (s, 1H), 8.38 (d, J=2.6 Hz, 1H), 8.25(td, J=2.2, 10.2 Hz, 1H), 8.19 (s, 1H), 7.34 (d, J=7.7 Hz, 1H),7.28-7.23 (m, 2H), 7.04-6.97 (m, 1H), 6.96-6.89 (m, 1H), 4.82-4.74 (m,1H), 4.59 (s, 1H), 3.39-3.31 (m, 1H), 3.27-3.21 (m, 1H), 2.93 (t, J=6.2Hz, 2H), 2.83 (dd, J=7.2, 15.1 Hz, 1H), 2.35 (dt, J=2.5, 6.4 Hz, 1H),2.25-2.13 (m, 1H), 1.38 (dd, J=2.3, 6.7 Hz, 6H); ES-LCMS m/z 441.3[M+H]⁺.

Example 30

Synthesis of I-31

Synthetic Scheme:

Step 1: Ethyl7-hydroxy-3-isopropyl-pyrazolo[1,5-a]pyrimidine-6-carboxylate

A mixture of diethyl 2-(ethoxymethylene)propanedioate (621.89 mg, 2.88mmol, 581.21 μL, 1 eq), 4-isopropyl-1H-pyrazol-5-amine (400 mg, 2.88mmol, 1 eq) in AcOH (3 mL) was degassed and purged with N₂ for 3 times.The mixture was stirred at 120° C. for 2 h under N₂ atmosphere. Thereaction mixture was cooled to 0° C. and filtered. The solid was washedwith ethanol and petroleum ether and dried under reduced pressure togive ethyl 7-hydroxy-3-isopropyl-pyrazolo[1,5-a]pyrimidine-6-carboxylate(700 mg, 2.65 mmol, 92.3% yield, 94.5% purity) as a white solid whichwas used in the next step without further purification. ¹H NMR (400 MHz,CD₃OD) δ ppm 8.55 (s, 1H), 7.91 (s, 1H), 4.34 (q, J=7.2 Hz, 2H), 3.11(m, 1H), 1.40-1.31 (m, 9H); ES-LCMS m/z 250.2 [M+H]⁺.

Step 2: 3-Isopropylpyrazolo[1,5-a]pyrimidin-7-ol

To a solution of ethyl7-hydroxy-3-isopropyl-pyrazolo[1,5-a]pyrimidine-6-carboxylate (300 mg,1.20 mmol, 1 eq) in H₂O (3 mL) was added hydrochloric acid (6.12 g,61.27 mmol, 6 mL, 36.5%, 50.90 eq). The mixture was stirred at 120° C.for 12 h. The reaction mixture was concentrated under reduced pressureto give 3-isopropylpyrazolo[1,5-a]pyrimidin-7-ol (200 mg, 902.92 μmol,75.0% yield, 80.0% purity) as a black brown solid which was used in thenext step without further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm8.12 (s, 1H), 7.94 (d, J=7.5 Hz, 1H), 5.98 (d, J=7.3 Hz, 1H), 2.91-2.78(m, 1H), 1.25 (d, J=6.8 Hz, 6H); ES-LCMS m/z 178.1 [M+H]⁺.

Step 3: 7-Chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine

A solution of 3-isopropylpyrazolo[1,5-a]pyrimidin-7-ol (100 mg, 564.33μmol, 1 eq) in POCl₃ (88.6 g, 577.83 mmol, 53.70 mL, 1023.93 eq) wasstirred at 120° C. for 3 h. The reaction mixture was concentrated underreduced pressure to give a residue which was diluted with ice, thenNaHCO₃ solid was added to above solution until pH to 8. The reactionmixture was extracted with EtOAc (30 mL×3). The combined organic layerswere dried over Na₂SO₄, filtered and concentrated under reduced pressureto give 7-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine (100 mg, 511.12μmol, 90.6% yield) as brown oil which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.34 (d, J=4.4 Hz,1H), 8.11 (s, 1H), 6.93 (d, J=4.4 Hz, 1H), 2.92-2.88 (m, 1H), 1.29 (d,J=6.8 Hz, 6H); ES-LCMS m/z 196.1 [M+H]⁺

Step 4:N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-31)

A mixture of 7-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine (75.20 mg,384.36 μmol, 1 eq), 2-(1H-indol-3-yl)ethanamine (73.90 mg, 461.24 μmol,1.2 eq), DIEA (496.76 mg, 3.84 mmol, 669.49 μL, 10 eq) in i-PrOH (2 mL)was degassed and purged with N₂ for 3 times. The mixture was stirred at55° C. for 3 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Syneri Max-RP C12 100*30 5 u; mobile phase:[water (0.05% HCl)-ACN]; B %: 15%-45%, 12 min), followed bylyophilization to yieldN-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(22.81 mg, 54.77 μmol, 14.3% yield, 94.2% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃CD) δ ppm 8.14 (s, 1H), 7.80 (d, J=7.1 Hz,1H), 7.44 (d, J=7.9 Hz, 1H), 7.26 (d, J=8.2 Hz, 1H), 7.15-7.06 (m, 1H),6.98 (t, J=7.5 Hz, 1H), 6.78 (t, J=7.2 Hz, 1H), 5.95 (d, J=7.3 Hz, 1H),3.93 (t, J=6.5 Hz, 2H), 3.21 (t, J=6.5 Hz, 2H), 3.10 (m, 1H), 1.32 (d,J=6.8 Hz, 6H); ES-LCMS m/z 320.0 [M+H]⁺.

Example 31

Synthesis of I-32a

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-(4,5,6,7-tetrahydro-1H-indazol-6-yl)pyrazolo[1,5-a]pyrimidin-7-amine

A solution of 4,5,6,7-tetrahydro-1H-indazol-6-amine (40 mg, 291.58 μmol,1 eq),7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(84.77 mg, 291.58 μmol, 1 eq), DIEA (376.85 mg, 2.92 mmol, 507.89 μL, 10eq) in i-PrOH (4 mL) was stirred at 120° C. for 3 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by flash silica gel chromatography (from PE/EtOAC=100/1 to1/1, TLC: PE/EtOAc=1/1, R_(f)=0.5) to yield5-(5-fluoro-3-pyridyl)-3-isopropyl-N-(4,5,6,7-tetrahydro-1H-indazol-6-yl)pyrazolo[1,5-a]pyrimidin-7-amine(90 mg, 218.42 μmol, 74.9% yield, 95.0% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 9.16 (t, J=1.4 Hz, 1H), 8.52 (d, J=2.6 Hz,1H), 8.37 (td, J=2.2, 9.9 Hz, 1H), 7.93 (s, 1H), 7.37 (s, 1H), 6.77 (s,1H), 4.42 (s, 1H), 3.48-3.31 (m, 2H), 2.91-2.84 (m, 1H), 2.84-2.76 (m,2H), 2.31-1.97 (m, 2H), 1.40 (d, J=6.8 Hz, 6H); ES-LCMS m/z 492.2[M+H]⁺.

Step 2:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[(6S)-4,5,6,7-tetrahydro-1H-indazol-6-yl]pyrazolo[1,5-a]pyrimidin-7-amine(I-32)

5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-(4,5,6,7-tetrahydro-1H-indazol-6-yl)pyrazolo[1,5-a]pyrimidin-7-amine(130 mg, 327.45 μmol, 1 eq) was separated by SFC (column: OD (250 mm*30mm, 5 um); mobile phase: [0.1% NH₃H₂O MeOH]; B %: 45%-45%, min) to givepeak 1 (t_(R)=1.465 min) and peak 2 (t_(R)=1.687 min). The reactionmixture (peak 1) was concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: PhenomenexGemini 150*25 mm*10 um; mobile phase: [water (0.05% HCl)-ACN]; B %:35%-65%, 10 min), followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-3-isopropyl-N-[(6S)-4,5,6,7-tetrahydro-1H-indazol-6-yl]pyrazolo[1,5-a]pyrimidin-7-amine(39.42 mg, 76.66 μmol, 23.4% yield, 97.4% purity, 3HCl) (EE=99.3%,t_(R)=1.465 min), [α]²⁶ _(D)=−27.146, C=0.106 g/100 mL, MeOH) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.10 (s, 1H), 8.85 (d, J=2.2Hz, 1H), 8.50 (d, J=8.8 Hz, 1H), 8.27 (s, 1H), 8.09 (s, 1H), 7.09 (s,1H), 4.76 (s, 1H), 3.50 (dd, J=5.3, 16.5 Hz, 1H), 3.39 (m J=6.8 Hz, 1H),3.19-3.11 (m, 1H), 2.93 (dd, J=4.4, 7.9 Hz, 2H), 2.41-2.30 (m, 1H),2.28-2.14 (m, 1H), 1.40 (d, J=6.8 Hz, 6H); ES-LCMS m/z 392.2 [M+H]⁺.

Example 32

Synthesis of I-33

Synthetic Scheme:

Step 1: tert-Butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate

To a solution of5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(650 mg, 1.75 mmol, 1 eq) in 1,4-dioxane (30 mL) was added DMAP (639.72mg, 5.24 mmol, 3 eq) and (Boc)₂O (1.14 g, 5.24 mmol, 1.20 mL, 3 eq). Themixture was stirred at 110° C. for 12 h. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby flash silica gel chromatography (from PE/EtOAc=100/1 to 3/1, TLC:PE/EtOAc=3/1, R_(f)=0.75) to give compound tert-butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(575 mg, 985.06 μmol, 56.4% yield, 98.1% purity) was obtained as yellowsolid. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.74 (t, J=1.5 Hz, 1H), 8.55 (d,J=2.9 Hz, 1H), 8.19 (d, J=2.2 Hz, 1H), 8.01 (d, J=7.3 Hz, 1H), 7.87-7.81(m, 1H), 7.47 (d, J=7.7 Hz, 1H), 7.27-7.23 (m, 2H), 7.20-7.14 (m, 1H),6.77 (d, J=2.4 Hz, 1H), 6.57 (s, 1H), 4.25 (t, J=6.8 Hz, 2H), 3.13 (t,J=6.7 Hz, 2H), 1.62 (s, 9H), 1.42 (s, 9H); ES-LCMS m/z 573.3 [M+H]⁺.

Step 2: tert-Butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)-3-iodo-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate

A mixture of tert-butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(250 mg, 428.29 μmol, 1 eq) and NIS (192.72 mg, 856.58 μmol, 2 eq) inDCM (10 mL) and MeCN (10 mL) was stirred at 25° C. for 12 h. TLC(PE/EtOAc=4/1, R_(f)=0.40) showed the starting material was consumedcompletely. The reaction mixture was quenched with aqueous Na₂S₂O₃ (10mL) and extracted with DCM (10 mL×3). The organic layer was dried overNa₂SO₄, filtered and concentrated under reduced pressure to dryness togive a residue which was purified by flash silica gel chromatography(from PE/EtOAc=20/1 to 5/1, TLC: PE/EtOAc=4/1, R_(f)=0.40). The desiredfraction was concentrated under reduced pressure to dryness to givetert-butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)-3-iodo-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(280 mg, 400.84 μmol, 93.6% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.76 (s, 1H), 8.57 (d, J=2.4 Hz, 1H), 8.18(s, 1H), 8.00 (br s, 1H), 7.96-7.91 (m, 1H), 7.45 (d, J=7.6 Hz, 1H),7.26 (br s, 2H), 7.20-7.15 (m, 1H), 6.58 (s, 1H), 4.23 (t, J=6.8 Hz,2H), 3.12 (t, J=6.8 Hz, 2H), 1.62 (s, 9H), 1.42 (s, 9H); ES-LCMS m/z699.2 [M+H]⁺.

Step 3: tert-Butyl3-[2-[[3-acetyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate

A mixture of tert-butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)-3-iodo-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(150 mg, 214.74 μmol, 1 eq), tributyl(1-ethoxyvinyl)stannane (310.21 mg,858.95 μmol, 289.92 μL, 4 eq) and Pd(dppf)Cl₂ (31.43 mg, 42.95 μmol, 0.2eq) in toluene (2 mL) was bubbled with N₂ for 2 min and then sealed. Thereaction mixture was irradiated under microwave (1 bar) at 100° C. for 2h. To the mixture was added aqueous KF (10 mL, 1 g/10 mL). The mixturewas stirred at 25° C. for 30 min and then extracted with EtOAc (10mL×3). The organic layer was dried over Na₂SO₄, filtered andconcentrated under reduced pressure to dryness to give a residue whichwas purified by flash silica gel chromatography (from PE/EtOAc=1/0 to2/1, TLC: PE/EtOAc=4/1, R_(f)=0.20). The desired fraction wasconcentrated under reduced pressure to dryness to give tert-butyl3-[2-[[3-acetyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate(80 mg, 124.17 μmol, 57.8% yield, 95.4% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.78 (s, 1H), 8.66 (d, J=2.0 Hz, 1H), 8.61(s, 1H), 7.96 (br s, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.42 (d, J=8.0 Hz,1H), 7.24-7.21 (m, 2H), 7.14 (t, J=7.2 Hz, 1H), 6.65 (s, 1H), 4.33-4.27(m, 2H), 3.14 (t, J=6.4 Hz, 2H), 2.84 (s, 3H), 1.59 (s, 9H), 1.43 (s,9H); ES-LCMS m/z 615.3 [M+H]⁺.

Step 4:1-[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone

A mixture of tert-butyl3-[2-[[3-acetyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate(80 mg, 124.17 μmol, 1 eq) and TFA (4.62 g, 40.52 mmol, 3 mL, 326.32 eq)in DCM (9 mL) was stirred at 25° C. for 30 min. TLC (PE/EtOAc=1/1,R_(f)=0.50) showed the starting material was consumed completely. Thereaction mixture was concentrated under reduced pressure at 30° C. todryness. The residue was dissolved in water (10 mL), basified withsaturated aqueous NaHCO₃ until pH=8 and extracted with EtOAc (15 mL×3).The organic layer was dried over Na₂SO₄, filtered and concentrated underreduced pressure to dryness to give1-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone(50 mg, 101.34 μmol, 81.6% yield, 84.0% purity) as a yellow solid whichwas used in the next step without further purification. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 10.81 (s, 1H), 9.16 (s, 1H), 8.71 (d, J=2.8 Hz, 1H), 8.62(t, J=6.4 Hz, 1H), 8.53 (s, 1H), 8.26-8.20 (m, 1H), 7.65 (d, J=7.6 Hz,1H), 7.28 (d, J=8.0 Hz, 1H), 7.20 (s, 1H), 7.08-7.03 (m, 1H), 7.01-6.97(m, 1H), 6.86 (s, 1H), 3.94-3.85 (m, 2H), 3.13 (t, J=6.8 Hz, 2H), 2.71(s, 3H); ES-LCMS m/z 415.1 [M+H]⁺.

Step 5:1-[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanol(I-33)

To a solution of1-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone(50 mg, 101.34 μmol, 1 eq) in MeOH (5 mL) was added NaBH₄ (38.34 mg,1.01 mmol, 10 eq) and then the mixture was stirred at 25° C. for 1 h.TLC (PE/EtOAc=1/1, R_(f)=0.40) showed the starting material was consumedcompletely. The reaction mixture was quenched with water (10 mL) andconcentrated under reduced pressure. The residue was extracted withEtOAc (15 mL×3). The organic layer was dried over Na₂SO₄, filtered andconcentrated under reduced pressure to dryness to give a residue whichwas purified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10um; mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %:36%-66%, 10 min). The desired fraction was lyophilized to give1-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanol(3.29 mg, 7.90 μmol, 7.8% yield, 100.0% purity) as a white solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.79 (s, 1H), 8.51 (d, J=2.8 Hz, 1H), 8.04 (br s,1H), 7.95 (s, 1H), 7.84 (d, J=8.4 Hz, 1H), 7.69 (d, J=7.6 Hz, 1H), 7.37(d, J=8.0 Hz, 1H), 7.22 (d, J=6.8 Hz, 2H), 7.07 (s, 1H), 6.56 (br s,1H), 6.09 (s, 1H), 5.35 (s, 1H), 3.89-3.83 (m, 2H), 3.27 (t, J=6.4 Hz,2H), 3.21 (d, J=3.6 Hz, 1H), 1.70 (d, J=6.4 Hz, 3H); ES-LCMS m/z 417.2[M+H]⁺.

Example 33

Synthesis of I-34

Synthetic Scheme:

Step 1:(3R)—N-[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-34)

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 171.13 μmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(31.87 mg, 171.13 μmol, 1 eq) and DIEA (22.12 mg, 171.13 μmol, 29.81 μL,1 eq) in i-PrOH (2 mL) was stirred at 80° C. for 12 h. The reactionmixture was concentrated under reduced pressure to dryness to give aresidue which was purified by preparative HPLC (column: Syneri Max-RPC12 100*30 5 u; mobile phase: [water (0.05% HCl)-ACN]; B %: 50%-70%, 12min). The desired fraction was lyophilized to give(3R)—N-[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(29.03 mg, 55.63 μmol, 32.5% yield, 100.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.76 (s, 1H), 9.25 (s, 1H), 8.67(d, J=2.8 Hz, 1H), 8.46 (td, J=2.0, 10.4 Hz, 1H), 8.30 (br s, 1H), 8.04(s, 1H), 7.32 (d, J=7.6 Hz, 1H), 7.24 (d, J=8.0 Hz, 1H), 7.03 (s, 1H),6.97 (t, J=6.8 Hz, 1H), 6.93-6.87 (m, 1H), 4.58-4.40 (m, 1H), 3.14-2.95(m, 2H), 2.94-2.78 (m, 2H), 2.29 (s, 3H), 2.15 (br s, 2H); ES-LCMS m/z413.2 [M+H]⁺.

Example 34

Synthesis of I-35

Synthetic Scheme:

Step 1:(3R)—N-[5-(5-Fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-35)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidine(35 mg, 102.58 μmol, 1 eq) and(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (21.02 mg, 112.83 μmol, 1.1eq) in i-PrOH (3 mL) was added DIEA (39.77 mg, 307.73 μmol, 53.60 μL,3.0 eq). The mixture was stirred at 60° C. for 3 h. The reaction mixturewas concentrated under reduced pressure to give a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um;mobile phase: [water (0.05% HCl)-ACN]; B %: 60%-90%, 10 min), followedby lyophilization to yield(3R)—N-[5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(28.79 mg, 49.45 umol, 48.2% yield, 98.9% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.36 (s, 1H), 8.87-8.79 (m, 2H),8.34 (s, 1H), 7.37 (d, J=7.7 Hz, 1H), 7.26 (d, J=8.2 Hz, 1H), 7.17 (s,1H), 7.06-6.99 (m, 1H), 6.98-6.91 (m, 1H), 4.57-4.46 (m, 1H), 3.33-3.31(m, 1H), 3.17-3.04 (m, 1H), 3.00-2.88 (m, 2H), 2.42-2.33 (m, 1H),2.31-2.19 (m, 1H); ES-LCMS m/z 467.2 [M+H]⁺.

Example 35

Synthesis of I-36

Synthetic Scheme:

Step 1:(3R)—N-[6-Chloro-2-(5-fluoro-3-pyridyl)pyrimidin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine

A mixture of 4,6-dichloro-2-(5-fluoro-3-pyridyl)pyrimidine (100 mg,389.26 μmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (79.75mg, 428.19 μmol, 1.1 eq), DIEA (150.93 mg, 1.17 mmol, 203.41 μL, 3 eq)in i-PrOH (3 mL) was degassed and purged with N₂ for 3 times. Themixture was stirred at 55° C. for 12 h under N₂ atmosphere. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by flash silica gel chromatography (from PE/EtOAC=100/1 to3/1, TLC: PE/EtOAc=3/1, R_(f)=0.4) to yield(3R)—N-[6-chloro-2-(5-fluoro-3-pyridyl)pyrimidin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(130 mg, 320.18 μmol, 82.3% yield, 97.0% purity) as a white solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.38 (s, 1H), 8.55 (d, J=2.9 Hz, 1H), 8.34(d, J=9.0 Hz, 1H), 7.91 (s, 1H), 7.47 (d, J=7.7 Hz, 1H), 7.34 (d, J=7.9Hz, 1H), 7.21-7.16 (m, 1H), 7.14-7.10 (m, 1H), 6.34 (s, 1H), 3.25 (dd,J=5.0, 15.5 Hz, 1H), 3.04-2.74 (m, 4H), 2.23 (s, 2H); ES-LCMS m/z 394.1[M+H]⁺.

Step 2:(3R)—N-[2-(5-Fluoro-3-pyridyl)-6-pyrrolidin-1-yl-pyrimidin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-36)

(3R)—N-[6-Chloro-2-(5-fluoro-3-pyridyl)pyrimidin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(60 mg, 147.77 μmol, 1 eq) and pyrrolidine (727.50 mg, 10.23 mmol,853.87 μL, 69.22 eq) were taken up into a microwave tube in i-PrOH (2mL). The sealed tube was heated at 135° C. for 3 h under microwave. Thereaction mixture was concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: Agela DurashellC18 150*25 5 u; mobile phase: [water (0.05% HCl)-ACN]; B %: 30%-60%, 12min), followed by lyophilization to yield(3R)—N-[2-(5-fluoro-3-pyridyl)-6-pyrrolidin-1-yl-pyrimidin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(42.07 mg, 78.06 μmol, 52.8% yield, 99.8% purity, 3HCl) as a brownsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.83 (s, 1H), 9.30 (s, 1H), 8.85(s, 1H), 8.67-8.48 (m, 1H), 8.31 (s, 1H), 7.35 (d, J=7.5 Hz, 1H), 7.26(d, J=7.9 Hz, 1H), 7.03-6.98 (m, 1H), 6.96-6.89 (m, 1H), 5.78 (s, 1H),4.29 (m, 1H), 3.75-3.60 (m, 3H), 3.51 (s, 1H), 3.10 (dd, J=4.6, 15.0 Hz,1H), 3.02-2.89 (m, 2H), 2.76-2.63 (m, 1H), 2.22-2.02 (m, 2H), 1.98 (s,4H); ES-LCMS m/z 429.3 [M+H]⁺.

Example 36

Synthesis of I-37a

Synthetic Scheme:

Step 1:5-(5-Bromo-3-pyridyl)-3-isopropyl-N-(5-methoxyindan-2-yl)pyrazolo[1,5-a]pyrimidin-7-amine

To a solution of5-(5-bromo-3-pyridyl)-7-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine(100.00 mg, 264.48 μmol, 1 eq) in i-PrOH (5 mL) was added5-methoxyindan-2-amine (52.81 mg, 264.48 μmol, 1 eq, HCl) and DIEA(341.82 mg, 2.64 mmol, 460.67 μL, 10 eq). The mixture was stirred at 90°C. for 12 h. The reaction mixture was concentrated under reducedpressure to give a residue which was purified by flash silica gelchromatography (from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1, R_(f)=0.31)to yield5-(5-bromo-3-pyridyl)-3-isopropyl-N-(5-methoxyindan-2-yl)pyrazolo[1,5-a]pyrimidin-7-amine(120 mg, 250.84 μmol, 94.8% yield, 100% purity) as yellow oil. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.18 (d, J=1.8 Hz, 1H), 8.74 (d, J=2.2 Hz, 1H),8.58 (t, J=2.1 Hz, 1H), 7.87 (s, 1H), 7.18 (d, J=8.4 Hz, 1H), 6.83 (s,1H), 6.79 (dd, J=2.5, 8.3 Hz, 1H), 6.58 (d, J=7.7 Hz, 1H), 6.37 (s, 1H),4.71-4.60 (m, 1H), 3.81 (s, 3H), 3.50 (m, 2H), 3.37 (t, J=6.9 Hz, 1H),3.08 (m, 2H), 1.42 (d, J=6.8 Hz, 6H); ES-LCMS m/z 478.1, 480.1 [M+H]⁺.

Step 2:5-[3-Isopropyl-7-[(5-methoxyindan-2-yl)amino]pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile

To a solution of5-(5-bromo-3-pyridyl)-3-isopropyl-N-(5-methoxyindan-2-yl)pyrazolo[1,5-a]pyrimidin-7-amine(120.0 mg, 250.84 μmol, 1 eq) in DMF (6 mL) was added Zn(CN)₂ (117.83mg, 1.00 mmol, 4 eq) and Pd(PPh₃)₄ (57.97 mg, 50.17 μmol, 0.2 eq) underN₂. The mixture was stirred at 85° C. for 32 h. The reaction mixture wasquenched by addition of water (30 mL), then extracted with EtOAc (30mL×3). The combined organic layers were washed with brine (10 mL), driedover Na₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1, R_(f)=0.55) to yield5-[3-isopropyl-7-[(5-methoxyindan-2-yl)amino]pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(80 mg, 188.46 μmol, 75.1% yield, 100% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.47 (d, J=2.2 Hz, 1H), 8.94 (d, J=2.0 Hz, 1H),8.72 (t, J=2.1 Hz, 1H), 7.89 (s, 1H), 7.18 (d, J=8.2 Hz, 1H), 6.88-6.76(m, 2H), 6.65 (d, J=7.5 Hz, 1H), 6.38 (s, 1H), 4.73-4.58 (m, 1H), 3.82(s, 3H), 3.51 (m, 2H), 3.36 (t, J=7.0 Hz, 1H), 3.09 (m, 2H), 1.42 (d,J=6.8 Hz, 6H); ES-LCMS m/z 425.2 [M+H]⁺.

Step 3:5-[7-[[(2S)-5-Hydroxyindan-2-yl]amino]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(I-37)

A mixture of5-[3-isopropyl-7-[(5-methoxyindan-2-yl)amino]pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(35.00 mg, 74.21 μmol, 1 eq) and pyridine; hydrochloride (428.76 mg,3.71 mmol, 50 eq) was stirred at 190° C. for 1 h under N₂. The reactionmixture was quenched by addition of sat. aq. NaHCO₃ (20 mL) slowly, thenextracted with EtOAc (20 mL×3). The combined organic layers were washedwith brine (10 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by preparative TLC(PE/EtOAc=1/1, TLC: PE/EtOAc=1/1, R_(f)=0.53) to yield5-[7-[(5-hydroxyindan-2-yl)amino]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(15 mg, 34.20 μmol, 46.0% yield, 93.6% purity) as a yellow solid (Note:Tatol 80 mg of target from two batches was used for SFC separation). Theracemate was separated by chiral SFC (AD (250 mm×30 mm, 10 μm); mobilephase: [0.1% NH₃H₂O IPA]; B %: 55%-55%, min; peak 1 (Rt=1.817) and peak2 (Rt=2.477)). The solution after separation was concentrated underreduced pressure to give a residue which was purified by preparativeHPLC (column: Phenomenex Gemini 150×25 mm×10 μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 50%-80%, 10 min) followed by lyophilization toyield5-[7-[[(2S)-5-hydroxyindan-2-yl]amino]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(19.05 mg, 39.41 μmol, 22.7% yield, 100% purity, 2HCl) (EE=100%,Rt=1.817 min, [α]²⁶ _(D)=3.608 (c=1.03 mg/mL, MeOH)) as a yellow solid.¹H NMR (400 MHz, CD₃OD) δ ppm 9.44 (d, J=2.2 Hz, 1H), 9.08 (d, J=2.0 Hz,1H), 8.84 (t, J=2.1 Hz, 1H), 8.09 (s, 1H), 7.07 (d, J=8.2 Hz, 1H), 6.88(s, 1H), 6.71 (s, 1H), 6.64 (dd, J=2.2, 8.2 Hz, 1H), 4.96-4.90 (m, 1H),3.50-3.41 (m, 2H), 3.38-3.33 (m, 1H), 3.19-3.03 (m, 2H), 1.40 (d, J=7.1Hz, 6H); ES-LCMS m/z 411.2 [M+H]⁺.

Example 37

Synthesis of I-38

Synthetic Scheme:

Step 1:(3R)—N-[5-(5-Bromo-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine

A mixture of5-(5-bromo-3-pyridyl)-7-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine(150 mg, 396.73 μmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(77.59 mg, 416.56 μmol, 1.05 eq) and DIEA (153.82 mg, 1.19 mmol, 207.31μL, 3 eq) in i-PrOH (10 mL) was degassed and purged with N₂ for 3 times.The mixture was stirred at 80° C. for 2 h under N₂ atmosphere. Thereaction mixture was concentrated under reduced pressure to give aresidue which was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1, R_(f)=0.34) to give(3R)—N-[5-(5-bromo-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(150 mg, 296.16 μmol, 74.6% yield, 99.0% purity) as a green solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.17 (d, J=1.8 Hz, 1H), 8.73 (d, J=2.0 Hz,1H), 8.60-8.55 (m, 1H), 7.90-7.87 (m, 1H), 7.47 (d, J=7.7 Hz, 1H), 7.34(d, J=8.2 Hz, 1H), 7.19 (br t, J=7.6 Hz, 1H), 7.15-7.10 (m, 1H), 6.58(br d, J=8.8 Hz, 1H), 6.40 (s, 1H), 4.32 (br s, 1H), 3.45-3.32 (m, 2H),3.06-2.90 (m, 3H), 2.44-2.23 (m, 2H), 1.43 (d, J=6.8 Hz, 6H); ES-LCMSm/z 501.1, 503.1 [M+H]⁺.

Step 2:5-[3-Isopropyl-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(I-38)

A mixture of(3R)—N-[5-(5-bromo-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(80 mg, 157.95 μmol, 1 eq), Pd(PPh₃)₄ (73.01 mg, 63.18 μmol, 0.4 eq) andZn(CN)₂ (74.19 mg, 631.81 μmol, 4 eq) in DMF (5 mL) was degassed andpurged with N₂ for 3 times. The mixture was stirred at 85° C. for 19 hunder N₂ atmosphere. The mixture was concentrated and water (80 mL) wasadded, extracted with EtOAc (50 mL×3). The combined organic layers weredried over Na₂SO₄, filtered and concentrated to the residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 μm;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 70%-100%,10 min) to yield product of5-[3-isopropyl-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(11.7 mg, 26.14 μmol, 16.6% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 10.78 (s, 1H), 9.66 (d, J=2.0 Hz, 1H),9.08-9.01 (m, 2H), 8.03 (s, 1H), 7.96 (d, J=9.5 Hz, 1H), 7.36 (d, J=7.7Hz, 1H), 7.28 (d, J=7.9 Hz, 1H), 7.06 (s, 1H), 7.01 (t, J=7.4 Hz, 1H),6.96-6.90 (m, 1H), 4.53-4.41 (m, 1H), 3.32-3.22 (m, 1H), 3.19-3.00 (m,2H), 2.97-2.81 (m, 2H), 2.23-2.13 (m, 2H), 1.38 (d, J=6.8 Hz, 6H);ES-LCMS m/z 448.2 [M+H]⁺.

Example 38

Synthesis of I-39

Synthetic Scheme:

Step 1: 3-Bromo-7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (50 mg, 168.39μmol, 1 eq) in DCM (2 mL) and ACN (4 mL) was added NBS (32.97 mg, 185.23μmol, 1.1 eq). The mixture was stirred at 30° C. for 1 h under N₂atmosphere. The reaction mixture was concentrated under reduced pressureto give 3-bromo-7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(55.16 mg, 168.41 μmol, 100.0% yield, 100% purity) as a yellow solidwhich was used in the next step without further purification. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.12 (s, 1H), 8.65 (d, J=2.6 Hz, 1H), 8.43-8.19(m, 2H), 7.51 (s, 1H); ES-LCMS m/z 327.0, 329.0 [M+H]⁺.

Step 2:3-Bromo-5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-39)

To a solution of3-bromo-7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (55.16mg, 168.41 μmol, 1 eq) in i-PrOH (4 mL) was added DIEA (65.30 mg, 505.22μmol, 88.00 μL, 3 eq) and 2-(1H-indol-3-yl)ethanamine (53.96 mg, 336.81μmol, 2 eq). The mixture was stirred at 60° C. for 5 h under N₂atmosphere. The reaction mixture was concentrated under reduced pressureto give a residue which was purified by preparative HPLC (column: AgelaDurashell C18 150×25×5μ; mobile phase: [water (0.05% HCl)-ACN]; B %:40%-70%, 12 min) followed by lyophilization to yield3-bromo-5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(65 mg, 115.93 μmol, 68.8% yield, 100% purity, 3HCl) as a yellow solid.¹H NMR (400 MHz, CD₃OD) δ ppm 8.71 (s, 1H), 8.55 (s, 1H), 8.18 (s, 1H),7.70 (d, J=8.8 Hz, 1H), 7.62 (d, J=8.0 Hz, 1H), 7.16 (d, J=8.3 Hz, 1H),7.05-6.91 (m, 3H), 5.94 (s, 1H), 3.98 (t, J=5.9 Hz, 2H), 3.22 (t, J=6.0Hz, 2H); ES-LCMS m/z 451.1, 453.1 [M+H]⁺.

Example 39

Synthesis of I-40

Synthetic Scheme:

Step 1:(3R)—N-[2-(5-Fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-40)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(60.00 mg, 205.68 μmol, 1 eq),(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (42.14 mg, 226.25 μmol, 1.1eq), DIEA (79.75 mg, 617.05 μmol, 107.48 μL, 3 eq) in i-PrOH (4 mL) wasdegassed and purged with N₂ for 3 times. The mixture was stirred at 55°C. for 3 h under N₂ atmosphere. The reaction mixture was concentratedunder reduced pressure to give a residue which was purified bypreparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um; mobilephase: [water (0.05% HCl)-ACN]; B %: 70%-100%, 10 min), followed bylyophilization to yield(3R)—N-[2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(42.07 mg, 75.45 μmol, 36.7% yield, 98.8% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.46 (s, 1H), 8.81 (d, J=9.3 Hz,1H), 8.76 (s, 1H), 8.12-7.92 (m, 1H), 7.37 (d, J=7.7 Hz, 1H), 7.27 (d,J=7.9 Hz, 1H), 7.08-7.00 (m, 1H), 6.99-6.90 (m, 1H), 3.34 (s, 1H),3.31-3.25 (m, 2H), 3.18-2.79 (m, 3H), 2.44-2.21 (m, 2H), 1.42 (d, J=7.1Hz, 6H); ES-LCMS m/z 442.2 [M+H]⁺.

Example 40

Synthesis of I-41

Synthetic Scheme:

Step 1:(3R)—N-[5-(5-Fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-41)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (50 mg, 166.91μmol, 1.0 eq) in i-PrOH (3 mL) was added DIEA (64.71 mg, 500.72 μmol,87.21 μL, 3.0 eq) and (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (37.30mg, 200.29 μmol, 1.2 eq). The mixture was stirred at 60° C. for 12 h.The reaction mixture was concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (HCl condition; column:Phenomenex Gemini 150×25 mm×10 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 38%-68%, 10 min). The desired fraction was lyophilizedto yield(3R)—N-[5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(61.30 mg, 120.71 μmol, 72.3% yield, 100.0% purity, 3 HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.01 (s, 1H), 8.77 (d, J=2.6Hz, 1H), 8.34-8.27 (m, 2H), 7.39 (d, J=7.7 Hz, 1H), 7.28 (d, J=7.9 Hz,1H), 7.13 (s, 1H), 7.09-7.01 (m, 1H), 7.00-6.93 (m, 1H), 6.66 (d, J=2.2Hz, 1H), 4.68 (s, 1H), 3.37-3.32 (m, 2H), 3.14-3.01 (m, 2H), 2.44-2.28(m, 2H); ES-LCMS m/z 339.2 [M+H]⁺.

Example 41

Synthesis of I-42a

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[(5S)-4,5,6,7-tetrahydro-1H-indazol-5-yl]pyrazolo[1,5-a]pyrimidin-7-amine(I-42)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(60 mg, 206.38 μmol, 1 eq) in i-PrOH (5 mL) was added DIEA (266.73 mg,2.06 mmol, 359.47 μL, 10 eq) and 4,5,6,7-tetrahydro-1H-indazol-5-amine(43.36 mg, 206.38 μmol, 1 eq, 2HCl). The mixture was stirred at 90° C.for 12 h. The reaction mixture was concentrated under reduced pressureto give a residue which was purified by flash silica gel chromatography(from PE/EtOAc=1/0 to 0/1, TLC: PE/EtOAc=1/1, R_(f)=0.16) to yield aproduct. The product was separated by chiral SFC (AD (250 mm×30 mm, 5μm); mobile phase: [0.1% NH₃H₂O IPA]; B %: 35%-35%, min; peak 1(R_(t)=5.512) and peak 2 (R_(t)=6.038)). The solution after separationwas concentrated under reduced pressure to give a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150×25 mm×10 μm;mobile phase: [water (0.05% HCl)-ACN]; B %: 35%-65%, 10 min) followed bylyophilization to yield5-(5-fluoro-3-pyridyl)-3-isopropyl-N-[(5S)-4,5,6,7-tetrahydro-1H-indazol-5-yl]pyrazolo[1,5-a]pyrimidin-7-amine(29.07 mg, 58.0 μmol, 24.1% yield, 100% purity, 3HCl) (EE=94.6%,R_(t)=5.512 min, [α]²⁶ _(D)=−18.785 (c 1.05 mg/mL, MeOH)) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.07 (s, 1H), 8.77 (d, J=2.4 Hz,1H), 8.39 (td, J=2.2, 9.2 Hz, 1H), 8.20 (s, 1H), 7.96 (s, 1H), 6.98 (s,1H), 4.57 (s, 1H), 3.40-3.34 (m, 1H), 3.26 (dd, J=5.2, 15.3 Hz, 1H),3.15-3.04 (m, 2H), 2.92 (dd, J=9.6, 15.3 Hz, 1H), 2.45-2.36 (m, 1H),2.31-2.19 (m, 1H), 1.41 (d, J=7.1 Hz, 6H); ES-LCMS m/z 392.2 [M+H]⁺.

Example 42

Synthesis of I-43

Synthetic Scheme:

Step 1: Methyl 3-(5-bromo-3-pyridyl)-3-oxo-propanoate & methyl(Z)-3-(5-bromo-3-pyridyl)-3-hydroxy-prop-2-enoate

To a mixture of 5-bromopyridine-3-carboxylic acid (5 g, 24.75 mmol, 1eq) and TEA (2.50 g, 24.75 mmol, 3.45 mL, 1 eq) in THE (100 mL) wasadded CDI (6.02 g, 37.13 mmol, 1.5 eq) in one portion at 30° C. underN₂. The mixture was stirred at 30° C. for 1 h. potassium;3-methoxy-3-oxo-propanoate (7.73 g, 49.50 mmol, 2 eq) and MgCl₂ (4.71 g,49.50 mmol, 2 eq) was added and the mixture was stirred at 30° C. for 16h. The mixture was adjusted pH to 5-6 with 3 NHCl, extracted with EtOAc(50 mL×3). The combined organic layers were dried over Na₂SO₄, filteredand concentrated under reduced pressure to give a residue which waspurified on silica gel column chromatography (from PE/EtOAc=1/0 to 10/3,TLC: PE/EtOAc=3/1, R_(f)=0.47) to give methyl3-(5-bromo-3-pyridyl)-3-oxo-propanoate (1.85 g, 6.45 mmol, 26.1% yield,90.0% purity) and methyl(Z)-3-(5-bromo-3-pyridyl)-3-hydroxy-prop-2-enoate (1.85 g, 6.45 mmol,26.1% yield, 90.0% purity) as a white solid. ¹H NMR (400 MHz, CDCl₃) δppm 12.46 (s, 1H), 9.05 (d, J=2.0 Hz, 1H), 8.90-8.87 (m, 2H), 8.75 (d,J=2.2 Hz, 1H), 8.39-8.36 (m, 1H), 8.21 (t, J=2.1 Hz, 1H), 5.72 (s, 1H),4.02 (s, 2H), 3.84 (s, 3H), 3.78 (s, 3H); ES-LCMS m/z 258.0, 260.0[M+H]⁺.

Step 2: 5-(5-Bromo-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol

A mixture of methyl 3-(5-bromo-3-pyridyl)-3-oxo-propanoate (1.11 g, 3.87mmol, 1 eq) and 1H-pyrazol-5-amine (386.37 mg, 4.65 mmol, 1.2 eq) inAcOH (5 mL) was stirred at 120° C. for 1 h. The reaction mixture wasconcentrated under reduced pressure to give5-(5-bromo-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol (1 g, 3.44 mmol,88.7% yield, crude purity) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δppm 10.31 (s, 1H), 9.00 (d, J=2.0 Hz, 1H), 8.88 (d, J=2.0 Hz, 1H), 8.52(t, J=2.1 Hz, 1H), 7.91 (d, J=2.0 Hz, 1H), 7.51 (s, 1H), 6.25-6.21 (m,2H); ES-LCMS m/z 293.0, 295.0 [M+H]⁺.

Step 3: 5-(5-Bromo-3-pyridyl)-7-chloro-pyrazolo[1,5-a]pyrimidine

A solution of 5-(5-bromo-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol (1 g,3.44 mmol, 1 eq) in POCl₃ (8 mL) was stirred at 100° C. for 3 h. Thereaction mixture was concentrated under reduced pressure to give aresidue which was purified by preparative-TLC (TLC: PE/EtOAc=3/1,R_(f)=0.65) to yield5-(5-bromo-3-pyridyl)-7-chloro-pyrazolo[1,5-a]pyrimidine (0.8997 g, 2.91mmol, 84.6% yield, 100.0% purity) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.18 (d, J=2.0 Hz, 1H), 8.81 (d, J=2.2 Hz, 1H), 8.62 (t,J=2.0 Hz, 1H), 8.30 (d, J=2.4 Hz, 1H), 7.44 (s, 1H), 6.91 (d, J=2.4 Hz,1H); ES-LCMS m/z 309.0, 311.0 [M+H]⁺.

Step 4:5-(5-Bromo-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine

To a mixture of 5-(5-bromo-3-pyridyl)-7-chloro-pyrazolo[1,5-a]pyrimidine(500 mg, 1.62 mmol, 1 eq) and 2-(1H-indol-3-yl)ethanamine (388.18 mg,2.42 mmol, 1.5 eq) in i-PrOH (20 mL) was added DIEA (626.28 mg, 4.85mmol, 844.04 μL, 3 eq) in one portion. The mixture was stirred at 80° C.for 2 h. The reaction mixture was concentrated under reduced pressure togive a residue which was purified on silica gel column chromatography(from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1, R_(f)=0.35) to yield5-(5-bromo-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(629 mg, 1.45 mmol, 89.9% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.87 (d, J=1.8 Hz, 1H), 8.71 (d, J=2.2 Hz,1H), 8.41-8.37 (m, 1H), 8.08 (br s, 1H), 8.01 (d, J=2.2 Hz, 1H), 7.68(d, J=7.5 Hz, 1H), 7.38 (d, J=8.2 Hz, 1H), 7.23-7.17 (m, 2H), 7.11 (s,1H), 6.61 (br s, 1H), 6.55 (d, J=2.2 Hz, 1H), 6.12 (s, 1H), 3.86 (q,J=6.2 Hz, 2H), 3.28 (t, J=6.6 Hz, 2H); ES-LCMS m/z 435.0, 437.0 [M+H]⁺.

Step 5:5-[7-[2-(1H-Indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(I-43)

A mixture of5-(5-bromo-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(150 mg, 346.18 μmol, 1 eq), Zn(CN)₂ (162.60 mg, 1.38 mmol, 87.89 μL, 4eq) and Pd(PPh₃)₄ (80.01 mg, 69.24 μmol, 0.2 eq) in DMF (5 mL) wasdegassed and purged with N₂ for 3 times. The mixture was stirred at 85°C. for 19 h under N₂ atmosphere. The mixture was concentrated andsaturated NaHCO₃ solution (10 mL) was added. The mixture was extractedwith EtOAc (50 mL×3). The combined organic layers were dried overNa₂SO₄, filtered and concentrated. To the crude product was added MeOH(30 mL) and stirred for 10 min. The suspension was filtered and solidwas collected, washed with PE/EtOAc (2/1, 30 mL×2), dried under vacuumto yield5-[7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(67.84 mg, 176.30 μmol, 50.9% yield, 98.6% purity) as a yellow solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 10.80 (br s, 1H), 9.46 (d, J=2.2 Hz, 1H),9.08 (d, J=2.0 Hz, 1H), 8.76 (t, J=2.1 Hz, 1H), 8.19 (br s, 1H), 8.14(d, J=2.4 Hz, 1H), 7.65 (d, J=7.9 Hz, 1H), 7.29 (d, J=7.9 Hz, 1H), 7.21(s, 1H), 7.09-6.97 (m, 2H), 6.70 (s, 1H), 6.52 (d, J=2.2 Hz, 1H), 3.85(d, J=6.0 Hz, 2H), 3.14 (t, J=7.3 Hz, 2H); ES-LCMS m/z 380.2 [M+H]⁺.

Example 43

Synthesis of I-44

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-indan-2-yl-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-44)

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(50 mg, 168.55 μmol, 1 eq), indan-2-amine (26.94 mg, 202.25 μmol, 1.2eq) and DIEA (65.35 mg, 505.64 μmol, 88.07 μL, 3 eq) in i-PrOH (5 mL)was stirred at 80° C. for 2 h. The mixture was concentrated underreduced pressure to give a residue which was purified by preparativeHPLC (column: Phenomenex Gemini 150*25 mm*10 μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 60%-90%, 10 min) to give5-(5-fluoro-3-pyridyl)-N-indan-2-yl-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(27.63 mg, 59.44 μmol, 35.3% yield, 99.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.32 (t, J=1.6 Hz, 1H), 8.66 (d,J=2.8 Hz, 1H), 8.50-8.46 (m, 1H), 8.16 (d, J=8.4 Hz, 1H), 8.01 (s, 1H),7.28-7.25 (m, 2H), 7.20-7.17 (m, 2H), 7.00 (s, 1H), 4.95-4.84 (m, 1H),3.43-3.38 (m, 2H), 3.26-3.22 (m, 1H), 3.20-3.14 (m, 2H), 1.37 (d, J=7.2Hz, 6H); ES-LCMS m/z 388.2 [M+H]⁺.

Example 44

Synthesis of I-45

Synthetic Scheme:

Step 1: 3-Isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidin-7-ol

A mixture of methyl 3-oxo-3-phenyl-propanoate (100 mg, 561.21 μmol, 1eq), 4-isopropyl-1H-pyrazol-5-amine (70.25 mg, 561.21 μmol, 1 eq) inAcOH (33.70 mg, 561.21 μmol, 32.10 μL, 1 eq) was degassed and purgedwith N₂ for 3 times. The mixture was stirred at 120° C. for 1 h under N₂atmosphere. The reaction mixture was concentrated under reduced pressureto yield 3-isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidin-7-ol (90 mg,355.31 μmol, 63.3% yield) as black brown oil which was used in the nextstep without further purification. ES-LCMS m/z 254.1 [M+H]⁺.

Step 2: 7-Chloro-3-isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidine

A solution of 3-isopropyl-5-phenyl-pyrazolo [1,5-a] pyrimidin-7-ol (90mg, 355.31 μmol, 1 eq) in POCl₃ (2 mL) was stirred at 110° C. for 3 h.The reaction mixture was concentrated under reduced pressure, dilutedwith DCM (10 mL×2), concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAC=100/1 to 10/1, TLC: PE/EtOAc=10/1, R_(f)=0.55) to yield7-chloro-3-isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidine (60 mg, 207.11μmol, 58.3% yield, 93.8% purity) as a yellow solid. ¹H NMR (400 MHz,CD₃OD) δ ppm 8.15-8.07 (m, 3H), 7.57-7.49 (m, 3H), 7.40 (s, 1H),3.37-3.48 (m, 1H), 1.45 (d, J=6.8 Hz, 6H); ES-LCMS m/z 272.1 [M+H]⁺.

Step 3:N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-45)

A mixture of 7-chloro-3-isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidine (60mg, 207.11 μmol, 1 eq), 2-(1H-indol-3-yl)ethanamine (49.77 mg, 310.66μmol, 1.5 eq) and DIEA (133.84 mg, 1.04 mmol, 180.37 μL, 5 eq) in i-PrOH(3 mL) was degassed and purged with N₂ for 3 times. The mixture wasstirred at 50° C. for 3 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Synergi C18 250*50 mm*10 um;mobile phase: [water (0.05% HCl)-ACN]; B %: 35%-65%, 10 min), followedby lyophilization to yieldN-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-5-phenyl-pyrazolo[1,5-a]pyrimidin-7-amine(38.50 mg, 81.29 μmol, 39.3% yield, 98.9% purity, 2HCl) as a gray solid.¹H NMR (400 MHz, CD₃OD) δ ppm 8.12 (s, 1H), 7.58-7.52 (m, 2H), 7.49-7.43(m, 2H), 7.23 (d, J=8.2 Hz, 1H), 7.20-7.15 (m, 2H), 7.07-7.02 (m, 2H),6.90-6.84 (m, 1H), 5.71 (s, 1H), 3.96 (t, J=5.8 Hz, 2H), 3.29-3.23 (m,1H), 3.23-3.19 (m, 2H), 1.33 (d, J=6.8 Hz, 6H); ES-LCMS m/z 396.2[M+H]⁺.

Example 45

Synthesis of I-46

Synthetic Scheme:

Step 1: 3-Isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol

A mixture of methyl 3-oxo-3-(3-pyridyl)propanoate (100 mg, 558.12 μmol,1 eq) and 4-isopropyl-1H-pyrazol-5-amine (69.86 mg, 558.12 μmol, 1 eq)in AcOH (2 mL) was stirred at 120° C. for 0.5 h. The reaction mixturewas concentrated under reduced pressure to give the crude product3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol (120 mg, 471.91μmol, 84.6% yield, crude purity) as brown oil which was used in the nextstep without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm9.47-8.95 (m, 1H), 8.80-8.70 (m, 1H), 8.30-8.20 (m, 1H), 7.87, 7.38 (m,1H), 7.60-7.50 (m, 1H), 7.08, 1.24 (d, J=6.8 Hz, 3H), 1.07 (dd, J=3.5,6.8 Hz, 3H); ES-LCMS m/z 255.1 [M+H]⁺.

Step 2: 7-Chloro-3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidine

A solution of 3-Isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol(120.00 mg, 471.91 μmol, 1 eq) in POCl₃ (4.95 g, 32.28 mmol, 3 mL, 68.41eq) was stirred at 110° C. for 3 h. The reaction mixture wasconcentrated under reduced pressure, diluted with DCM (20 mL) which waspurified by preparative-TLC (PE/EtOAc=3/1, TLC: PE/EtOAc=3/1,R_(f)=0.40) to give7-chloro-3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidine (58 mg,212.66 μmol, 45.1% yield, 100.0% purity) as a green solid. ¹H NMR (400MHz, CDCl₃) δ ppm 9.32 (br s, 1H), 8.75 (br s, 1H), 8.45 (d, J=6.4 Hz,1H), 8.12 (br s, 1H), 7.52-7.44 (m, 1H), 7.43-7.39 (m, 1H), 3.45-3.40(m, 1H), 1.51-1.41 (m, 6H); ES-LCMS m/z 273.1 [M+H]⁺.

Step 3:N-[2-(1H-Indol-3-yl)ethyl]-3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-amine(I-46)

To a solution of7-chloro-3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidine (40 mg,146.66 μmol, 1 eq) and 2-(1H-indol-3-yl)ethanamine (35.25 mg, 220.00μmol, 1.5 eq) in i-PrOH (5 mL) was added DIEA (56.87 mg, 439.99 μmol,76.64 μL, 3 eq). The mixture was stirred at 60° C. for 3 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 25%-55%, 10 min) toyieldN-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-amine(31.67 mg, 60.73 μmol, 41.4% yield, 97.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.84 (dd, J=1.2, 5.4 Hz, 1H), 8.74(d, J=1.8 Hz, 1H), 8.11 (s, 1H), 8.04-7.98 (m, 1H), 7.91 (dd, J=5.2, 7.8Hz, 1H), 7.57 (d, J=7.9 Hz, 1H), 7.14 (d, J=7.9 Hz, 1H), 7.01-6.95 (m,2H), 6.91-6.86 (m, 1H), 5.81 (s, 1H), 4.00-3.94 (m, 2H), 3.24-3.16 (m,3H), 1.32 (d, J=6.8 Hz, 6H); ES-LCMS m/z 397.2 [M+H]⁺.

Example 46

Synthesis of I-47

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-7-ol

To a solution of methyl(Z)-3-(5-fluoro-3-pyridyl)-3-hydroxy-prop-2-enoate (100.00 mg, 486.91μmol, 1 eq) in AcOH (2 mL) was added4-(trifluoromethyl)-1H-pyrazol-5-amine (91.32 mg, 486.91 μmol, 1 eq,HCl). The mixture was stirred at 120° C. for 2 h. The reaction mixturewas concentrated under reduced pressure to give the crude product5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-7-ol(100 mg, 335.35 μmol, 68.9% yield, crude) as a yellow solid which wasused in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.00 (d, J=1.3 Hz, 1H), 8.87-8.79 (m, 2H), 8.35 (s, 1H),8.23-8.11 (m, 2H); ES-LCMS m/z 299.0 [M+H]⁺.

Step 2:7-Chloro-5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidine

A solution of5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-7-ol(100 mg, 335.35 μmol, 1 eq) in POCl₃ (18.85 g, 122.94 mmol, 11.42 mL,366.59 eq) was stirred at 110° C. for 3 h. The reaction mixture wasconcentrated under reduced pressure to remove solvent. The residue wasdiluted with DCM (20 mL×2) and concentrated under reduced pressure togive a residue which was purified on silica gel column chromatography(from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1, R_(f)=0.64) to give theproduct compound7-chloro-5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidine(34 mg, 80.53 μmol, 24.0% yield, 75.0% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.13 (s, 1H), 8.67 (s, 1H), 8.48 (s, 1H), 8.30(d, J=6.4 Hz, 1H), 7.65 (s, 1H); ES-LCMS m/z 317.1, 319.0 [M+H]⁺.

Step 3:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-7-amine(I-47)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidine(30 mg, 71.06 μmol, 1 eq) and 2-(1H-indol-3-yl)ethanamine (17.08 mg,106.59 μmol, 1.5 eq) in i-PrOH (5 mL) was added DIEA (27.55 mg, 213.18μmol, 37.13 μL, 3.0 eq). The mixture was stirred at 60° C. for 3 h. Thereaction mixture was concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: PhenomenexGemini 150*25 mm*10 μm; mobile phase: [water (0.05% ammonia hydroxidev/v)-ACN]; B %: 52%-82%, 10 min) followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-7-amine(24.84 mg, 45.09 μmol, 63.5% yield, 99.8% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.77 (br s, 1H), 8.67 (s, 1H), 8.28(s, 1H), 7.91 (d, J=8.2 Hz, 1H), 7.71-7.63 (m, 1H), 7.12-7.06 (m, 1H),7.04-6.96 (m, 2H), 6.90 (s, 1H), 6.02 (s, 1H), 3.90 (t, J=6.1 Hz, 2H),3.16 (t, J=6.1 Hz, 2H); ES-LCMS m/z 441.2 [M+H]⁺.

Example 47

Synthesis of I-48

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-9-isopropyl-N-(5-methoxyindan-2-yl)purin-6-amine

To a solution of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine (50mg, 164.55 μmol, 1 eq) in i-PrOH (5 mL) was added DIEA (170.13 mg, 1.32mmol, 229.29 μL, 8 eq) and 5-methoxyindan-2-amine (33.60 mg, 168.27μmol, 1.02 eq, HCl) under N₂. The mixture was stirred at 60° C. for 12h. The reaction mixture was concentrated under reduced pressure to givecrude2-(5-fluoro-3-pyridyl)-9-isopropyl-N-(5-methoxyindan-2-yl)purin-6-amine(60 mg, 102.80 μmol, 62.4% yield, 71.7% purity) as black brown oil whichwas used in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 11.16 (s, 1H), 9.54 (s, 1H), 8.51 (d, J=2.9 Hz, 1H), 8.45(d, J=9.0 Hz, 1H), 7.83 (s, 1H), 7.17 (d, J=7.9 Hz, 1H), 6.83 (s, 1H),6.77 (dd, J=2.5, 8.3 Hz, 1H), 4.92 (m, 1H), 4.04 (td, J=6.1, 12.3 Hz,1H), 3.84-3.79 (m, 3H), 3.48 (dt, J=7.3, 16.5 Hz, 2H), 3.05-2.92 (m,2H), 1.22 (d, J=6.2 Hz, 6H); ES-LCMS m/z 419.2 [M+H]⁺.

Step 2:2-(5-Fluoro-3-pyridyl)-9-isopropyl-N-(5-methoxyindan-2-yl)purin-6-amine(I-48)

A solution of2-(5-fluoro-3-pyridyl)-9-isopropyl-N-(5-methoxyindan-2-yl)purin-6-amine(60 mg, 102.80 μmol, 1 eq) in HBr (5 mL, 60% in water) was stirred at120° C. for 2 h. The reaction mixture was concentrated under reducedpressure to give a residue which was purified by preparative HPLC(column: Phenomenex Gemini C18 250×50 mm×10 μm; mobile phase:[water(0.05% HCl)-ACN]; B %: 40%-70%, 10 min) followed by lyophilizationto yield2-[[2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-yl]amino]indan-5-ol(21.36 mg, 44.75 μmol, 43.5% yield, 100% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.58 (s, 1H), 9.22 (s, 1H), 8.98(d, J=8.4 Hz, 1H), 8.90 (s, 1H), 7.09 (d, J=8.2 Hz, 1H), 6.73 (s, 1H),6.65 (dd, J=2.4, 8.2 Hz, 1H), 5.24 (s, 1H), 5.16 (td, J=6.8, 13.6 Hz,1H), 3.46 (ddd, J=7.2, 12.7, 15.8 Hz, 2H), 3.02 (dt, J=4.5, 14.9 Hz,2H), 1.75 (d, J=6.1 Hz, 6H); ES-LCMS m/z 405.1 [M+H]⁺

Example 48

Synthesis of I-49

Synthetic Scheme:

Step 1:4-[2-[[5-(5-Fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenol(I-49)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (50 mg, 164.89μmol, 1 eq) in i-PrOH (5 mL) was added DIEA (170.49 mg, 1.32 mmol,229.77 μL, 8 eq) and 4-(2-aminoethyl)phenol (33.93 mg, 247.34 μmol, 1.50eq) under N₂. The mixture was stirred at 80° C. for 12 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by preparative HPLC (column: Phenomenex Gemini 150×25 mm×10μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 20%-50%, 10 min),followed by lyophilization to yield4-[2-[[5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenol(46.38 mg, 109.83 μmol, 66.6% yield, 100% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.85-8.75 (m, 2H), 8.27 (d, J=2.2Hz, 1H), 8.09 (td, J=2.3, 9.2 Hz, 1H), 7.07 (d, J=8.6 Hz, 2H), 6.65-6.56(m, 3H), 6.46 (s, 1H), 3.97 (t, J=6.6 Hz, 2H), 3.00 (t, J=6.5 Hz, 2H);ES-LCMS m/z 350.2 [M+H]⁺.

Example 49

Synthesis of I-50

Synthetic Scheme:

Step 1: (6E)-6-Hydroxyimino-5H-cyclopenta[f][1,3]benzodioxol-7-one

A suspension of 5,6-dihydrocyclopenta[f][1,3]benzodioxol-7-one (0.5 g,2.84 mmol, 1 eq) in MeOH (45 mL) was heated to 45° C., then isopentylnitrite (539.40 mg, 4.60 mmol, 0.62 mL, 1.62 eq) and con. HCl (12 M inwater, 0.47 mL, 1.99 eq) were added. The mixture was stirred at 45° C.for 1.5 h. The mixture was filtered, the cake was washed with cold MeOH(5 mL×2), dried in vacuo to give a product (300 mg). The filtrate wasconcentrate under reduced pressure to give a residue which was addedMeOH (5 mL), then was filtered, dried in vacuo to give a product (200mg). Compound (6E)-6-hydroxyimino-5H-cyclopenta[f][1,3]benzodioxol-7-one(500 mg, 2.44 mmol, 85.8% yield, 100% purity) was obtained as a yellowsolid which was used in the next step without further purification. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 12.47 (s, 1H), 7.14 (d, J=10.4 Hz, 2H),6.18 (s, 2H), 3.64 (s, 2H); ES-LCMS m/z 206.1 [M+H]⁺.

Step 2: 6,7-Dihydro-5H-cyclopenta[f][1,3]benzodioxol-6-amine

To a solution of(6E)-6-hydroxyimino-5H-cyclopenta[f][1,3]benzodioxol-7-one (500 mg, 2.44mmol, 1 eq) in AcOH (25 mL) and con. H₂SO₄ (0.3 mL) was added Pd/C (0.13g, 10%). The mixture was stirred at 25° C. for 12 h under H₂ (30 psi).The mixture was filtered, the filtrate was concentrated under reducedpressure to give a residue which was added water (50 mL), adjusted pH to10-11 with 2 N aq. NaOH, extracted with DCM (30 mL×3). The combinedorganic layers were washed with brine (20 mL), dried over Na₂SO₄,filtered and concentrated under reduced pressure to give6,7-dihydro-5H-cyclopenta[f][1,3]benzodioxol-6-amine (250 mg, 1.13 mmol,46.3% yield, 80% purity) as a yellow solid which was used in the nextstep without further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 6.68(s, 2H), 5.90 (s, 2H), 3.91-3.76 (m, 1H), 3.09 (dd, J=6.8, 15.3 Hz, 2H),2.58 (dd, J=4.9, 15.4 Hz, 2H); ES-LCMS m/z No correct mass was found.

Step 3:N-(6,7-Dihydro-5H-cyclopenta[f][1,3]benzodioxol-6-yl)-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-amine(I-50)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (60 mg, 197.87μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (127.87 mg, 989.37 μmol,172.33 μL, 5 eq) and6,7-dihydro-5H-cyclopenta[f][1,3]benzodioxol-6-amine (65.74 mg, 296.81μmol, 1.5 eq). The mixture was stirred at 90° C. for 12 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by preparative HPLC (column: Phenomenex Gemini C18 250×50mm×10 μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 40%-70%, 10 min)followed by lyophilization to yieldN-(6,7-dihydro-5H-cyclopenta[f][1,3]benzodioxol-6-yl)-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-amine(21.53 mg, 46.43 μmol, 23.4% yield, 99.7% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.02 (t, J=1.3 Hz, 1H), 8.81 (d,J=2.6 Hz, 1H), 8.34-8.29 (m, 1H), 8.28 (d, J=2.2 Hz, 1H), 7.05 (s, 1H),6.76 (s, 2H), 6.66 (d, J=2.2 Hz, 1H), 5.94-5.90 (m, 2H), 5.08 (t, J=6.8Hz, 1H), 3.46 (dd, J=7.7, 15.4 Hz, 2H), 3.18 (dd, J=6.3, 15.5 Hz, 2H);ES-LCMS m/z 390.1 [M+H]⁺.

Example 50

Synthesis of I-51

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-(5-methoxyindan-2-yl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (35mg, 133.25 μmol, 1.0 eq) in i-PrOH (4 mL) was added DIEA (86.10 mg,666.24 μmol, 116.04 μL, 5.0 eq) and 5-methoxyindan-2-amine (30 mg,150.24 μmol, 1.13 eq, HCl salt). The mixture was stirred at 60° C. for12 h. The reaction mixture was concentrated under reduced pressure togive5-(5-fluoro-3-pyridyl)-N-(5-methoxyindan-2-yl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(50 mg, 113.24 μmol, 85.0% yield, 88.2% purity) was obtained as yellowoil which was used in the next step without further purification. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.07 (s, 1H), 8.55 (d, J=2.6 Hz, 1H), 8.21 (d,J=9.9 Hz, 1H), 7.86 (s, 1H), 7.18 (d, J=8.8 Hz, 1H), 6.87-6.75 (m, 2H),6.60 (m, 1H), 6.39 (s, 1H), 4.66 (s, 1H), 3.81 (s, 3H), 3.70-3.64 (m,4H), 2.39 (s, 3H); ES-LCMS m/z 390.2 [M+H]⁺.

Step 2:2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(I-51)

5-(5-Fluoro-3-pyridyl)-N-(5-methoxyindan-2-yl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(50 mg, 113.24 μmol, 1 eq) was added into HBr (5 mL, 60% in water) andthe mixture was stirred at 120° C. for 2 h. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (HCl condition; column: Phenomenex Gemini 150×25mm×10 um; mobile phase: [water(0.05% HCl)-ACN]; B %: 35%-65%, 10 min).The desired fraction was lyophilized to yield2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(15.05 mg, 33.46 μmol, 29.5% yield, 99.6% purity, 2 HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.03 (s, 1H), 8.75 (d, J=2.5Hz, 1H), 8.31 (td, J=2.3, 9.3 Hz, 1H), 8.07 (s, 1H), 7.08 (d, J=8.3 Hz,1H), 6.89 (s, 1H), 6.72 (s, 1H), 6.68-6.62 (m, 1H), 4.99-4.95 (m, 1H),3.50-3.41 (m, 2H), 3.21-3.10 (m, 2H), 2.37 (s, 3H); ES-LCMS m/z 376.1[M+H]⁺.

Example 51

Synthesis of I-52

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-(5-methoxyindan-2-yl)pyrazolo[1,5-a]pyrimidin-7-amine

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (50 mg, 164.89μmol, 1.0 eq) in i-PrOH (6 mL) was added DIEA (106.56 mg, 824.47 μmol,143.61 μL, 5.0 eq) and 5-methoxyindan-2-amine (35 mg, 175.28 μmol, 1.06eq, HCl salt). The mixture was stirred at 60° C. for 12 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas used in the next step without further purification. Compound5-(5-fluoro-3-pyridyl)-N-(5-methoxyindan-2-yl)pyrazolo[1,5-a]pyrimidin-7-amine(60 mg, 139.21 μmol, 84.4% yield, 87.1% purity) was obtained as a yellowoil. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.97 (s, 1H), 8.49 (d, J=2.9 Hz, 1H),8.09 (td, J=2.3, 9.5 Hz, 1H), 7.94 (d, J=2.2 Hz, 1H), 7.11 (d, J=8.4 Hz,1H), 6.79-6.69 (m, 2H), 6.51 (d, J=2.2 Hz, 1H), 6.37 (s, 1H), 4.63-4.55(m, 1H), 3.77-3.68 (m, 3H), 3.59 (dt, J=3.0, 6.6 Hz, 4H); ES-LCMS m/z376.2 [M+H]⁺.

Step 2:2-[[5-(5-Fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol

5-(5-Fluoro-3-pyridyl)-N-(5-methoxyindan-2-yl)pyrazolo[1,5-a]pyrimidin-7-amine(60 mg, 139.21 μmol, 1 eq) was added into HBr (6 mL, 60% in water) andthe mixture was stirred at 120° C. for 2 h. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (HCl condition; column: Phenomenex Gemini 150×25mm×10 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 30%-60%, 10 min).The desired fraction was lyophilized to yield2-[[5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(32.33 mg, 72.95 μmol, 52.4% yield, 98.0% purity, 2 HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.01 (s, 1H), 8.82 (s, 1H),8.34-8.26 (m, 2H), 7.12-7.05 (m, 2H), 6.73 (s, 1H), 6.69-6.63 (m, 2H),5.06 (s, 1H), 3.47 (td, J=8.3, 15.9 Hz, 2H), 3.27-3.15 (m, 2H); ES-LCMSm/z 362.1 [M+H]⁺.

Example 52

Synthesis of I-53

Synthetic Scheme:

Step 1:5-[4-Chloro-6-(cyclopentoxy)pyrimidin-2-yl]pyridine-3-carbonitrile

To a solution of cyclopentanol (20.84 mg, 241.96 umol, 21.96 μL, 1.5 eq)in THE (3 mL) was added NaH (9.68 mg, 241.96 μmol, 60% in mineral oil,1.5 eq). The mixture was stirred at 0° C. for 30 min.5-(4,6-dichloropyrimidin-2-yl)pyridine-3-carbonitrile (50 mg, 161.31μmol, 1 eq) was added into the above solution and the mixture wasstirred at 28° C. for 12 h. The reaction mixture was quenched byaddition NH₄Cl (2 mL) and water (1 mL), then extracted with EtOAc (20mL×3). The combined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedby preparative TLC (PE/EtOAc=13/1, TLC: PE/EtOAc=13/1, R_(f)=0.54) toyield 5-[4-chloro-6-(cyclopentoxy) pyrimidin-2-yl]pyridine-3-carbonitrile (35 mg, 108.23 μmol, 67.1% yield, 93.0% purity)as a white solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.70 (d, J=2.2 Hz, 1H),9.04 (d, J=2.0 Hz, 1H), 9.01 (t, J=2.1 Hz, 1H), 6.91 (s, 1H), 5.72-5.67(m, J=3.1, 6.0 Hz, 1H), 2.15-2.08 (m, 2H), 1.95-1.88 (m, 2H), 1.87-1.83(m, 2H), 1.77-1.70 (m, 2H); ES-LCMS m/z 301.1 [M+H]⁺.

Step 2:5-[4-(Cyclopentoxy)-6-[2-(1H-indol-3-yl)ethylamino]pyrimidin-2-yl]pyridine-3-carbonitrile(I-53)

5-[4-Chloro-6-(cyclopentoxy)pyrimidin-2-yl]pyridine-3-carbonitrile (60mg, 185.54 μmol, 1 eq), 2-(1H-indol-3-yl)ethanamine (74.32 mg, 463.85μmol, 2.5 eq) and DIEA (23.98 mg, 185.54 μmol, 32.32 uL, 1 eq) weretaken up into a microwave tube in i-PrOH (4 mL). The sealed tube washeated at 135° C. for 3 h under microwave. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Synergi C18 250*50 mm*10 um;mobile phase: [water(0.05% HCl)-ACN]; B %: 60%-90%, 10 min), followed bylyophilization to yield5-[4-(cyclopentoxy)-6-[2-(1H-indol-3-yl)ethylamino]pyrimidin-2-yl]pyridine-3-carbonitrile(21.28 mg, 39.58 μmol, 21.3% yield, 99.3% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD3OD) δ ppm 10.81 (br s, 1H), 9.61 (s, 1H),9.10 (d, J=2.0 Hz, 1H), 8.87 (s, 1H), 7.56 (d, J=7.9 Hz, 1H), 7.46 (brs, 1H), 7.31 (d, J=8.2 Hz, 1H), 7.21-7.15 (m, 1H), 7.07-7.01 (m, 1H),7.00-6.94 (m, 1H), 5.72 (s, 1H), 5.39 (br s, 1H), 3.68 (br s, 2H), 2.96(t, J=7.4 Hz, 2H), 1.96 (d, J=6.0 Hz, 2H), 1.74-1.64 (m, 4H), 1.62-1.52(m, 2H); ES-LCMS m/z 425.2 [M+H]⁺.

Example 53

Synthesis of I-54

Synthetic Scheme:

Step 1: 5-(4,6-Dichloropyrimidin-2-yl)pyridine-3-carbonitrile

A mixture of 4,6-dichloro-2-iodo-pyrimidine (813.84 mg, 2.91 mmol, 1eq), (5-cyano-3-pyridyl)boronic acid (409.00 mg, 2.76 mmol, 0.95 eq),Pd(dppf)Cl₂ (212.96 mg, 291.04 μmol, 0.1 eq), Na₂CO₃ (925.42 mg, 8.73mmol, 3.0 eq) and water (2.4 mL) in 1,4-dioxane (12 mL) was degassed andpurged with N₂ for 3 times, then the mixture was stirred at 80° C. for 1h under N₂ atmosphere. The reaction mixture was diluted with EtOAc (50mL) and filtered through a pad of celite. The filtrate was concentratedunder reduced pressure to give a residue which was purified by flashsilica gel chromatography (from PE/EtOAC=100/1 to 5/1, TLC:PE/EtOAc=5/1, R_(f)=0.7) to yield5-(4,6-dichloropyrimidin-2-yl)pyridine-3-carbonitrile (180 mg, 580.71μmol, 20.0% yield, 81.0% purity) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.81 (d, J=2.4 Hz, 1H), 9.03 (d, J=2.0 Hz, 1H), 8.99 (d,J=2.0 Hz, 1H), 7.44 (s, 1H); ES-LCMS m/z 250.9, 252.9 [M+H]⁺.

Step 2: 5-(4-Chloro-6-isopropoxy-pyrimidin-2-yl)pyridine-3-carbonitrile

To a solution of i-PrOH (9.69 mg, 161.31 μmol, 12.35 μL, 1 eq) in THF (3mL) was added NaH (6.45 mg, 161.31 μmol, 60%, 1 eq). The mixture wasstirred at 0° C. for 30 min.5-(4,6-dichloropyrimidin-2-yl)pyridine-3-carbonitrile (50 mg, 161.31μmol, 1 eq) was added into the above solution. The mixture was stirredat 28° C. for 12 h. The reaction mixture was concentrated under reducedpressure to give a residue which was diluted with NH₄Cl (30 mL) andextracted with EtOAc (30 mL×3). The combined organic layers were driedover Na₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified by preparative TLC (PE/EtOAc=10/1, TLC:PE/EtOAc=10/1, R_(f)=0.55) to yield5-(4-chloro-6-isopropoxy-pyrimidin-2-yl) pyridine-3-carbonitrile (25 mg,87.37 μmol, 54.2% yield, 96.0% purity) as a white solid. ¹H NMR (400MHz, CD₃OD) δ ppm 9.70 (s, 1H), 9.05 (d, J=2.0 Hz, 1H), 9.01 (t, J=2.1Hz, 1H), 6.90 (s, 1H), 5.61 (td, J=6.3, 12.4 Hz, 1H), 1.44 (d, J=6.2 Hz,6H); ES-LCMS m/z 275.1 [M+H]⁺.

Step 3:5-[4-[2-(1H-indol-3-yl)ethylamino]-6-isopropoxy-pyrimidin-2-yl]pyridine-3-carbonitrile(I-54)

5-(4-chloro-6-isopropoxy-pyrimidin-2-yl) pyridine-3-carbonitrile (25 mg,91.01 μmol, 1 eq), 2-(1H-indol-3-yl)ethanamine (21.87 mg, 136.51 μmol,1.5 eq) and DIEA (35.29 mg, 273.02 μmol, 47.56 μL, 3 eq) were taken upinto a microwave tube in i-PrOH (3 mL). The sealed tube was heated at135° C. for 5 h under microwave. The reaction mixture was concentratedunder reduced pressure to give a residue which was purified bypreparative HPLC (column: Phenomenex Synergi C18 250*50 mm*10 um; mobilephase: [water(0.05% HCl)-ACN]; B %: 55%-85%, 10 min), followed bylyophilization to yield5-[4-[2-(1H-indol-3-yl)ethylamino]-6-isopropoxy-pyrimidin-2-yl]pyridine-3-carbonitrile(16.5 mg, 32.17 μmol, 35.3% yield, 99.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.84 (s, 1H), 9.62 (s, 1H), 9.12(d, J=2.0 Hz, 1H), 8.90 (s, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.33 (d, J=7.9Hz, 1H), 7.20 (s, 1H), 7.09-7.02 (m, 1H), 7.01-6.95 (m, 1H), 5.74 (s,1H), 5.41-5.30 (m, 1H), 3.71 (m, 2H), 2.97 (t, J=7.3 Hz, 2H), 1.31 (d,J=6.2 Hz, 6H); ES-LCMS m/z 399.2 [M+H]⁺.

Example 54

Synthesis of I-55

Synthetic Scheme:

Step 1:[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]-(1-piperidyl)methanone(I-55)

To a solution of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (50 mg, 120.07 μmol, 1 eq) in pyridine (2 mL) was added piperidine(15.34 mg, 180.11 μmol, 17.79 μL, 1.5 eq) and T₃P (152.82 mg, 240.15μmol, 142.82 μL, 50%, 2 eq). The mixture was stirred at 25° C. for 12 hunder N₂ atmosphere. The reaction mixture was concentrated under reducedpressure to give a residue which was diluted with EtOAc (10 mL) andextracted with EtOAc (30 mL×3). The combined organic layers were washedwith brine (10 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by preparativeHPLC (column: Phenomenex Gemini 150*25 mm*10 um; mobile phase:[water(0.05% HCl)-ACN]; B %: 42%-72%, 10 min), followed bylyophilization to yield[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]-(1-piperidyl)methanone(12.74 mg, 21.49 μmol, 17.9% yield, 100.0% purity, 3HCl) as a whitesolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.72-8.68 (m, 1H), 8.57 (s, 1H),8.34 (s, 1H), 7.71 (td J=2.4, 8.9 Hz, 1H), 7.60 (d, J=7.9 Hz, 1H), 7.14(d, J=7.9 Hz, 1H), 7.02-6.96 (m, 2H), 6.94-6.89 (m, 1H), 6.03 (s, 1H),4.00 (t, J=6.0 Hz, 2H), 3.71 (m, 4H), 3.23 (t, J=6.1 Hz, 2H), 1.75 (m,2H), 1.68 (m, 4H); ES-LCMS m/z 484.2 [M+H]⁺.

Example 55

Synthesis of I-56

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]-N,N-dimethyl-pyrazolo[1,5-a]pyrimidine-3-carboxamide(I-56)

A mixture of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (40 mg, 96.06 μmol, 1 eq), N-methylmethanamine (23.50 mg, 288.18μmol, 3 eq, HCl) and T₃P (611.29 mg, 960.60 μmol, 571.30 μL, 50% inEtOAc, 10 eq) in pyridine (5 mL) was stirred at 30° C. for 12 h. Themixture was concentrated under reduced pressure to give a residue whichwas diluted with EtOAc (20 mL) and water (20 mL), extracted with EtOAc(20 mL×3), dried over Na₂SO₄, filtered and concentrated under reducedpressure to give a residue which was purified by preparative HPLC(column: Phenomenex Gemini 150*25 mm*10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 33%-63%, 10 min) to give5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]-N,N-dimethyl-pyrazolo[1,5-a]pyrimidine-3-carboxamide(16.67 mg, 29.67 μmol, 30.9% yield, 98.4% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) (ppm 10.82 (br s, 1H), 9.12 (s, 1H),8.69 (d, J=3.2 Hz, 1H), 8.43 (t, J=6.0 Hz, 1H), 8.31 (s, 1H), 8.21 (d,J=10.0 Hz, 1H), 7.63 (d, J=7.6 Hz, 1H), 7.29 (d, J=8.0 Hz, 1H), 7.23 (d,J=1.6 Hz, 1H), 7.07-7.02 (m, 1H), 7.00-6.97 (m, 1H), 6.78 (s, 1H), 3.87(q, J=6.8 Hz, 2H), 3.16-3.03 (m, 8H); ES-LCMS m/z 444.2 [M+H]+.

Example 56

Synthesis of I-57

Synthetic Scheme:

Step 1:1-[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone(I-57)

To a solution of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]-N-methoxy-N-methyl-pyrazolo[1,5-a]pyrimidine-3-carboxamide(50 mg, 96.85 μmol, 1 eq) in THE (10 mL) was added MeMgBr (3 M in Et₂O,4.68 mL, 145.10 eq) dropwise at 25° C. Then the mixture was stirred at25° C. for 0.5 h. The mixture was quenched with sat.NH₄Cl (20 mL),extracted with EtOAc (20 mL×3), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedby preparative TLC (SiO₂, PE/EA=1/1, R_(f)=0.56), then purified bypreparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um; mobilephase: [water(0.05% HCl)-ACN]; B %: 40%-70%, 10 min) to give1-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone(17.52 mg, 32.98 μmol, 34.1% yield, 98.6% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.81 (br s, 1H), 9.16 (s, 1H),8.72 (d, J=2.8 Hz, 1H), 8.62 (t, J=6.4 Hz, 1H), 8.53 (s, 1H), 8.23 (d,J=10.0 Hz, 1H), 7.65 (d, J=8.0 Hz, 1H), 7.28 (d, J=8.0 Hz, 1H), 7.20 (d,J=2.0 Hz, 1H), 7.06-7.03 (m, 1H), 7.02-6.99 (m, 1H), 6.87 (s, 1H), 3.89(q, J=6.8 Hz, 2H), 3.14 (t, J=7.2 Hz, 2H), 2.72 (s, 3H); ES-LCMS m/z415.2 [M+H]⁺.

Example 57

Synthesis of I-58

Synthetic Scheme:

Step 1: tert-Butyl4-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carbonyl]piperazine-1-carboxylate

A mixture of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (50 mg, 120.07 μmol, 1 eq), tert-butylpiperazine-1-carboxylate (26.84 mg, 144.09 μmol, 1.2 eq) and T₃P (382.06mg, 600.37 μmol, 357.06 uL, 50%, 5 eq) in pyridine (1 mL) was stirred at25° C. for 16 h. The reaction mixture was quenched with water (5 mL) andextracted with EtOAc (10 mL×3). The organic layer was dried over Na₂SO₄,filtered and concentrated under reduced pressure to dryness to givetert-butyl4-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carbonyl]piperazine-1-carboxylate(100 mg, crude) as a brown solid which was used in the next stepdirectly without further purification.

Step 2:[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]-piperazin-1-yl-methanone(I-58)

To solution of tert-butyl4-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carbonyl]piperazine-1-carboxylate (100 mg,171.04 μmol, 1 eq) in MeOH (3 mL) was added HCl/MeOH (4 M, 3 mL) andthen the mixture was stirred at 25° C. for 2 h. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um; mobilephase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 20%-50%, 10 min).The desired fraction was lyophilized to give[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]-piperazin-1-yl-methanone(19.42 mg, 39.99 μmol, 23.4% yield, 99.8% purity) as an off-white solid.¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.80 (s, 1H), 9.12 (s, 1H), 8.69 (d,J=2.8 Hz, 1H), 8.41 (t, J=6.0 Hz, 1H), 8.32 (s, 1H), 8.23-8.16 (m, 1H),7.63 (d, J=8.4 Hz, 1H), 7.29 (d, J=8.0 Hz, 1H), 7.22 (d, J=2.4 Hz, 1H),7.05 (t, J=7.2 Hz, 1H), 7.00-6.95 (m, 1H), 6.77 (s, 1H), 3.87 (q, J=6.8Hz, 2H), 3.54 (m, 4H), 3.13 (t, J=7.2 Hz, 2H), 2.77-2.73 (m, 4H);ES-LCMS m/z 485.2 [M+H]⁺.

Example 58

Synthesis of I-59

Synthetic Scheme:

Step 1: 4,6-Dichloro-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine

To a suspension of 2-(1H-indol-3-yl)ethanamine (666.18 mg, 4.16 mmol,1.05 eq) in dry THE (10 mL) was added NaH (223.55 mg, 5.59 mmol, 60% inmineral oil, 1.41 eq) under ice bath and N₂ atmosphere. After beingstirred for 30 min, the suspension was cooled to −60° C. and a solutionof 4,6-dichloro-2-methylsulfonyl-pyrimidine (900 mg, 3.96 mmol, 1.0 eq)in dry THE (10 mL) was added dropwise and kept the temperature below−55° C. The resulting mixture was stirred for 1 h at −55° C. Thereaction mixture was poured into water (100 mL) slowly and extractedwith EtOAc (50 mL×3). The combined organic layers were washed withbrine, dried over Na₂SO₄ and concentrated to give the residue which waspurified by flash silica gel chromatography (from PE/EtOAc=100/1 to 2/1,TLC: PE/EtOAc=3/1, R_(f)=0.45) to yield4,6-dichloro-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine (840 mg, 2.68mmol, 67.6% yield, 98.0% purity) as a white solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 8.04 (s, 1H), 7.68 (d, J=7.9 Hz, 1H), 7.36 (d, J=7.9 Hz,1H), 7.23 (dt, J=1.1, 7.6 Hz, 1H), 7.19-7.14 (m, 1H), 6.98 (d, J=2.2 Hz,1H), 6.59 (s, 1H), 5.74 (s, 1H), 3.83-3.73 (m, 2H), 3.07 (t, J=6.8 Hz,2H); ES-LCMS m/z 307.0, 309.0 [M+H]⁺.

Step 2:4-Chloro-6-(cyclopentoxy)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine

To a solution of NaH (30.63 mg, 765.68 μmol, 60% in mineral oil, 1.2 eq)in THE (3 mL) was added cyclopentanol (57.71 mg, 669.97 μmol, 60.81 μL,1.05 eq). The mixture was stirred at 0° C. for 30 min.4,6-dichloro-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine (200 mg, 638.07μmol, 1 eq) was added into the above solution and the mixture wasstirred at 15° C. for 12 h. The reaction mixture was concentrated underreduced pressure to give4-chloro-6-(cyclopentoxy)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine(170 mg, 309.66 μmol, 48.5% yield, 65.0% purity) as a yellow solid whichwas used in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 8.03 (s, 1H), 7.66 (t, J=7.5 Hz, 1H), 7.39 (d, J=8.2 Hz,1H), 7.25-7.19 (m, 1H), 7.18-7.10 (m, 1H), 7.08-7.02 (m, 1H), 6.60 (s,1H), 5.98 (s, 1H), 5.31 (m, 1H), 3.80-3.72 (m, 2H), 3.11-3.04 (m, 2H),2.01-1.54 (m, 8H); ES-LCMS m/z 357.1, 358.1 [M+H]⁺.

Step 3:4-(Cyclopentoxy)-6-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine(I-59)

4-Chloro-6-(cyclopentoxy)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine(170 mg, 304.89 μmol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (128.88mg, 914.67 μmol, 3 eq), Cs₂CO₃ (298.02 mg, 914.67 μmol, 3.0 eq) andPd(dppf)Cl₂ (22.31 mg, 30.49 μmol, 0.1 eq) were taken up into amicrowave tube in 1,4-dioxane (6 mL) and water (1.2 mL). The sealed tubewas heated at 80° C. for 30 min under microwave. The reaction mixturewas diluted with EtOAc (100 mL) and filtered through a pad of celite.The filtrate was concentrated under reduced pressure to give a residuewhich was purified by flash silica gel chromatography (fromPE/EtOAc=100/1 to 2/1, TLC: PE/EtOAc=3/1, R_(f)=0.40), then re-purifiedby preparative HPLC (HCl condition, column: Phenomenex Gemini 150×25mm×10 um; mobile phase: [water(0.05% HCl)-ACN]; B %: 48%-78%, 10 min).The desired fraction was lyophilized to yield4-(cyclopentoxy)-6-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine(25.91 mg, 48.46 μmol, 15.9% yield, 98.5% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.13 (s, 1H), 8.88 (s, 1H), 8.64(s, 1H), 8.40 (d, J=7.3 Hz, 1H), 8.15 (s, 1H), 7.63 (d, J=7.8 Hz, 1H),7.38 (d, J=8.0 Hz, 1H), 7.25 (s, 1H), 7.20 (t, J=7.4 Hz, 1H), 7.15-7.09(m, 1H), 6.32 (s, 1H), 5.27 (s, 1H), 3.85 (q, J=6.5 Hz, 2H), 3.17 (t,J=6.8 Hz, 2H), 1.94 (d, J=5.8 Hz, 2H), 1.84-1.74 (m, 4H), 1.70-1.62 (m,2H); ES-LCMS m/z 418.2 [M+H]⁺.

Example 59

Synthesis of I-60

Step 1:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-piperazin-1-yl-pyrimidin-4-amine

A mixture of6-chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine(60 mg, 161.50 μmol, 1 eq), piperazine (139.11 mg, 1.61 mmol, 10 eq) andDIEA (104.36 mg, 807.49 μmol, 140.65 μL, 5 eq) in i-PrOH (3 mL) wassealed and irradiated under microwave (4 bar) at 150° C. for 2 h. Thereaction mixture was concentrated under reduced pressure to dryness togive a residue which was purified by preparative HPLC (column:Phenomenex Gemini C18 250*50 mm*10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 8%-38%, 10 min). The desired fraction was lyophilized togive2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-piperazin-1-yl-pyrimidin-4-amine(60.34 mg, 106.30 μmol, 65.8% yield, 99.2% purity, 4HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.05 (s, 1H), 8.77 (d, J=2.8 Hz,1H), 8.33 (d, J=9.2 Hz, 1H), 7.64 (d, J=8.0 Hz, 1H), 7.33 (d, J=7.6 Hz,1H), 7.14 (s, 1H), 7.12-7.07 (m, 1H), 7.06-7.01 (m, 1H), 5.58 (br s,1H), 3.86-3.72 (m, 6H), 3.27-3.24 (m, 4H), 3.14 (t, J=6.0 Hz, 2H);ES-LCMS m/z 418.1 [M+H]⁺.

Example 60

Synthesis of I-61

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]-N-methyl-pyrazolo[1,5-a]pyrimidine-3-carboxamide(I-61)

To a solution of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (60 mg, 144.09 μmol, 1 eq) in anhydrous DCM (5 mL) was added DIEA(55.87 mg, 432.27 μmol, 75.29 μL, 3 eq), HATU (136.97 mg, 360.22 μmol,2.5 eq) and methanamine hydrochloride (19.46 mg, 288.18 μmol, 2 eq). Themixture was stirred at 25° C. for 12 h. H₂O (10 mL) was added, themixture was extracted with DCM (10 mL×3). The combined organic layerswere washed with brine (10 mL), dried over anhydrous Na₂SO₄, filteredand concentrated to afford the crude product which was purified bypreparative HPLC (MeCN/H₂O as eluents, acidic condition, Instrument:Phenomenex Gemini C18 250*50 mm*10 um/Mobile phase: water(0.05%HCl)-ACN/Gradient: B from 35% to 65% in 10 min/Flow rate: 25 mL/min)followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]-N-methyl-pyrazolo[1,5-a]pyrimidine-3-carboxamide(19.25 mg, 35.73 μmol, 24.79% yield, 100% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.73 (d, J=12.5 Hz, 2H), 8.47 (s,1H), 7.94 (d, J=9.0 Hz, 1H), 7.70 (dd, J=3.0, 6.0 Hz, 1H), 7.15 (dd,J=2.9, 6.1 Hz, 1H), 7.07-7.02 (m, 2H), 6.97 (s, 1H), 6.07 (s, 1H), 3.97(t, J=6.0 Hz, 2H), 3.23 (t, J=6.1 Hz, 2H), 3.04-2.99 (m, 3H); ES-LCMSm/z 452.1 [M+Na]⁺.

Example 61

Synthesis of I-62

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-(1-piperidyl)pyrimidin-4-amine(I-62)

6-Chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine(60.61 mg, 163.13 μmol, 1.0 eq), piperidine (862.20 mg, 10.13 mmol, 1mL, 62.07 eq) and DIEA (63.25 mg, 489.39 μmol, 85.24 μL, 3.0 eq) weretaken up into a microwave tube in i-PrOH (3 mL). The sealed tube washeated at 150° C. for 3 h under microwave. The reaction mixture wasconcentrated under reduced pressure to give the residue which waspurified by preparative HPLC (HCl condition; column: Phenomenex GeminiC18 250×50 mm×10 um; mobile phase: [water (0.05% HCl)-ACN]; B %:30%-60%, 10 min) and the desired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-(1-piperidyl)pyrimidin-4-amine(33.57 mg, 63.84 μmol, 39.1% yield, 100.0% purity, 3HCl salt) as ayellow solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 10.92 (s, 1H), 9.29 (s, 1H),8.83 (s, 1H), 8.52 (d, J=6.8 Hz, 1H), 7.98 (s, 1H), 7.59 (d, J=7.7 Hz,1H), 7.40-7.25 (m, 2H), 7.16-6.91 (m, 2H), 5.77 (s, 1H), 4.17 (s, 4H),3.63 (d, J=6.6 Hz, 2H), 3.02 (t, J=6.7 Hz, 2H), 1.75-1.47 (m, 6H);ES-LCMS m/z 417.2 [M+H]⁺.

Example 62

Synthesis of I-63

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-N6-[2-(1H-indol-3-yl)ethyl]-N4,N4-dimethyl-pyrimidine-4,6-diamine(I-63)

6-Chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine(80.81 mg, 217.51 umol, 1.0 eq), N-methylmethanamine (177.36 mg, 2.18mmol, 10 eq, HCl salt) and DIEA (168.66 mg, 1.31 mmol, 227.31 μL, 6.0eq) in i-PrOH (3 mL) were taken up into a microwave tube. The sealedtube was heated at 150° C. for 3 h under microwave. The reaction mixturewas concentrated under reduced pressure to give a residue which waspurified by preparative HPLC (HCl condition; column: Phenomenex Gemini150×25 mm×10 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 20%-50%, 10min) and the desired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N6-[2-(1H-indol-3-yl)ethyl]-N4,N4-dimethyl-pyrimidine-4,6-diamine(28.13 mg, 56.54 μmol, 26.0% yield, 97.7% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.93 (s, 1H), 9.31 (s, 1H), 8.85(d, J=2.0 Hz, 1H), 8.55 (d, J=8.8 Hz, 1H), 8.09 (s, 1H), 7.60 (d, J=7.7Hz, 1H), 7.42-7.25 (m, 2H), 7.10-7.04 (m, 1H), 7.02-6.96 (m, 1H), 5.59(s, 1H), 4.04 (s, 6H), 3.62 (t, J=6.7 Hz, 2H), 3.03 (t, J=6.7 Hz, 2H);ES-LCMS m/z 377.2 [M+H]⁺.

Example 63

Synthesis of I-64

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-morpholino-pyrimidin-4-amine(I-64)

To a solution of6-chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine(60 mg, 161.50 μmol, 1 eq) in i-PrOH (1.5 mL) was added DIEA (208.72 mg,1.61 mmol, 281.29 μL, 10 eq), morpholine (281.39 mg, 3.23 mmol, 284.24μL, 20 eq). The mixture was stirred at 150° C. for 1.5 h on microwave.The reaction mixture was concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: PhenomenexGemini 150×25 mm×10 μm; mobile phase: [water(0.05% HCl)-ACN]; B %:25%-55%, 10 min) followed by lyophilization to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-morpholino-pyrimidin-4-amine(53.33 mg, 101.03 μmol, 62.5% yield, 100% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.87 (s, 1H), 9.30 (s, 1H),8.86-8.32 (m, 2H), 7.59 (d, J=7.9 Hz, 1H), 7.35 (d, J=7.9 Hz, 1H), 7.25(s, 1H), 7.11-7.04 (m, 1H), 7.03-6.94 (m, 1H), 5.70 (s, 1H), 3.78-3.60(m, 11H), 3.01 (t, J=7.1 Hz, 2H); ES-LCMS m/z 419.2 [M+H]⁺.

Example 64

Synthesis of I-65

Synthetic Scheme:

Step 1:[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]-morpholino-methanone(I-65)

To a solution of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (40 mg, 96.06 μmol, 1 eq) in pyridine (3 mL) was added T₃P (122.26mg, 192.12 μmol, 114.26 μL, 50%, 2 eq) and morpholine (16.74 mg, 192.12μmol, 16.91 μL, 2.0 eq). The mixture was stirred at 25° C. for 12 h. Thereaction mixture was quenched by addition of water (50 mL), thenextracted with EtOAc (30 mL×3). The combined organic layers were washedwith brine (10 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by preparativeHPLC (column: Phenomenex Gemini C18 250×50 mm×10 μm; mobile phase:[water(0.05% HCl)-ACN]; B %: 30%-60%, 10 min) followed by lyophilizationto yield[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]-morpholino-methanone(19.50 mg, 32.66 μmol, 34.0% yield, 99.6% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.71-8.59 (m, 2H), 8.34 (s, 1H),7.73 (d, J=9.3 Hz, 1H), 7.63 (d, J=7.7 Hz, 1H), 7.14 (d, J=7.7 Hz, 1H),7.05-6.90 (m, 3H), 6.06 (s, 1H), 3.97 (t, J=6.1 Hz, 2H), 3.75 (s, 8H),3.22 (t, J=5.8 Hz, 2H); ES-LCMS m/z 486.2 [M+H]⁺.

Example 65

Synthesis of I-66

Step 1: 7-Fluoro-2-methyl-3-methylsulfanyl-1H-indole

2-Fluoroaniline (0.9 g, 8.10 mmol, 782.61 μL, 1 eq) and1-methylsulfanylpropan-2-one (371.24 mg, 3.56 mmol, 0.44 eq) wereinitially introduced into n-butyl acetate (4 ml) and cooled to −30° C.under N₂. A solution of sulfuryl chloride (437.28 mg, 3.24 mmol, 323.91μL, 0.4 eq) in n-butyl acetate (4 ml) was added dropwise. The mixturewas stirred for 2 h at −30° C. The reaction mixture was quenched byaddition of water (50 mL) at 0° C., then extracted with EtOAc (30 mL×3).The combined organic layers were washed with brine (10 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 10/1, TLC: PE/EtOAc=20/1, R_(f)=0.34) to yield a product7-fluoro-2-methyl-3-methylsulfanyl-1H-indole (1 g, 4.56 mmol, 56.3%yield, 89.0% purity) as red-brown oil. ¹H NMR (400 MHz, CDCl₃) δ ppm8.31 (s, 1H), 7.45 (d, J=7.7 Hz, 1H), 7.07 (dt, J=4.7, 7.9 Hz, 1H), 6.89(dd, J=7.8, 10.9 Hz, 1H), 2.56 (s, 3H), 2.27 (s, 3H); ES-LCMS m/z 195.9[M+H]⁺.

Step 2: 7-Fluoro-2-methyl-1H-indole

To a solution of 7-fluoro-2-methyl-3-methylsulfanyl-1H-indole (737.08mg, 3.36 mmol, 1 eq) in TFA (8 mL) was added 2-sulfanylbenzoic acid(1.30 g, 8.40 mmol, 2.5 eq). The mixture was stirred at 25° C. for 2 h.TLC (PE/EtOAc=20/1, R_(f)=0.40) showed the starting material wasconsumed completely and a new spot formed. The reaction mixture wasquenched by addition of water (100 mL), then adjusted to Ph to 10 by 1Naq. NaOH, extracted with EtOAc (50 mL×3). The combined organic layerswere washed with brine (30 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedby flash silica gel chromatography (from PE/EtOAc=1/0 to 10/1, TLC:PE/EtOAc=20/1, R_(f)=0.40) to yield 7-fluoro-2-methyl-1H-indole (400 mg,2.09 mmol, 62.3% yield, 78.0% purity) as yellow oil. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 11.33 (s, 1H), 7.20 (d, J=7.5 Hz, 1H), 6.92-6.74 (m, 2H),6.19 (s, 1H), 2.38 (s, 3H); ES-LCMS m/z No correct mass was found.

Step 3: 7-Fluoro-2-methyl-1H-indole-3-carbaldehyde

To a solution of DMF (10 mL) was added POCl₃ (481.07 mg, 3.14 mmol,291.56 μL, 2.0 eq) dropwise at −20° C. over a period of 10 min under N₂.After 1 h, 7-fluoro-2-methyl-1H-indole (300 mg, 1.57 mmol, 1 eq) in DMF(2 mL) was added to the above solution during which the temperature wasmaintained below −20° C. The reaction mixture was stirred at 15° C. for1 h. The reaction mixture was quenched by addition of aq. NaHCO₃ (50mL), then extracted with EtOAc (30 mL×3). The combined organic layerswere washed with brine (10 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedby flash silica gel chromatography (from PE/EtOAc=1/0 to 3/1, TLC:PE/EtOAc=3/1, R_(f)=0.40) to yield7-fluoro-2-methyl-1H-indole-3-carbaldehyde (200 mg, 936.93 μmol, 59.7%yield, 83.0% purity) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm12.44 (s, 1H), 10.08 (s, 1H), 7.85 (d, J=7.7 Hz, 1H), 7.13 (dt, J=5.1,7.8 Hz, 1H), 7.02 (dd, J=7.9, 11.2 Hz, 1H), 2.70 (s, 3H); ES-LCMS m/z177.9 [M+H]⁺.

Step 4: 7-Fluoro-2-methyl-3-[(E)-2-nitrovinyl]-1H-indole

To a solution of 7-fluoro-2-methyl-1H-indole-3-carbaldehyde (200 mg,936.93 μmol, 1 eq) in nitromethane (8 mL) was added NH₄OAc (216.66 mg,2.81 mmol, 3 eq). The mixture was stirred at 110° C. for 12 h. Thereaction mixture was concentrated under reduced pressure to give aresidue. The residue was dissolved in EtOAc (50 mL), washed with water(10 mL), brine (10 mL), dried over Na₂SO₄, filtered and concentratedunder reduced pressure to give a crude which was purified by flashsilica gel chromatography (from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1,R_(f)=0.61) to yield 7-fluoro-2-methyl-3-[(E)-2-nitrovinyl]-1H-indole(150 mg, 619.89 μmol, 66.2% yield, 91.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.82 (br s, 1H), 8.32 (d, J=13.5 Hz, 1H),7.76 (d, J=13.5 Hz, 1H), 7.46 (d, J=7.9 Hz, 1H), 7.25-7.16 (m, 1H), 7.01(dd, J=8.2, 10.6 Hz, 1H), 2.67 (s, 3H); ES-LCMS m/z 220.9 [M+H]⁺.

Step 5: 2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethanamine

To a solution of 7-fluoro-2-methyl-3-[(E)-2-nitrovinyl]-1H-indole (150mg, 619.89 μmol, 1 eq) in THF (10 mL) was added dropwise LAH (1 M inTHF, 3.10 mL, 5 eq) at 0° C. After addition, the mixture was stirred at80° C. for 2 h. The reaction mixture was diluted with THE (50 mL),quenched by addition of water (0.05 mL), aq. NaOH (0.05 mL, 10% inwater) and water (0.15 mL) in sequence at 0° C. After being stirred for30 min, the mixture was filtered through celite, the filtrate wasconcentrated under reduced pressure to give a crude2-(7-fluoro-2-methyl-1H-indol-3-yl)ethanamine (100 mg, 436.97 μmol,70.5% yield, 84.0% purity) as yellow oil which was used in the next stepdirectly without further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm7.23 (d, J=7.8 Hz, 1H), 6.93-6.82 (m, 1H), 6.72 (dd, J=7.9, 11.4 Hz,1H), 2.90-2.80 (m, 4H), 2.39 (s, 3H); ES-LCMS m/z 193.2 [M+H]⁺.

Step 6:N-[2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo(I-66)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(40.82 mg, 137.59 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (53.35 mg,412.76 μmol, 71.89 μL, 3.0 eq) and2-(7-fluoro-2-methyl-1H-indol-3-yl)ethanamine (37.78 mg, 165.11 μmol,1.2 eq). The mixture was stirred at 60° C. for 12 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by preparative HPLC (column: Gemini 150×25×5 μm; mobilephase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 70%-100%, 10 min)followed by lyophilization to yieldN-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(20.36 mg, 45.36 μmol, 33.0% yield, 99.5% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 8.58 (s, 1H), 8.45 (d, J=2.6 Hz, 1H), 7.89(s, 1H), 7.64-7.57 (m, 1H), 7.37 (d, J=7.9 Hz, 1H), 6.96 (dt, J=4.6, 7.8Hz, 1H), 6.69 (dd, J=7.8, 11.6 Hz, 1H), 5.76 (s, 1H), 3.83 (t, J=6.2 Hz,2H), 3.29-3.22 (m, 1H), 3.13 (t, J=6.1 Hz, 2H), 2.11 (s, 3H), 1.38 (d,J=7.1 Hz, 6H); ES-LCMS m/z 447.2 [M+H]⁺.

Example 66

Synthesis of I-67

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-(4-methylpiperazin-1-yl)pyrimidin-4-amine(I-67)

To a solution of6-chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine(60 mg, 161.50 μmol, 1 eq) and 1-methylpiperazine (24.26 mg, 242.25μmol, 26.87 uL, 1.5 eq) in i-PrOH (5 mL) was added DIEA (62.62 mg,484.49 μmol, 84.39 μL, 3.0 eq). The mixture was stirred at 110° C. for16 h. LC-MS showed 83% of starting material was remained and 17% ofdesired compound was detected. The mixture was taken up into a microwavetube. 1-methylpiperazine (500 mg, 5 mmol) was added. The mixture waspurged with N₂ for 1 min. The sealed tube was heated at 150° C. for 3 hunder microwave. The reaction mixture was concentrated under reducedpressure to give a residue which was purified by preparative HPLC(column: Phenomenex Gemini 150*25 mm*10 um; mobile phase: [water (0.05%ammonia hydroxide v/v)-ACN]; B %: 42%-72%, 10 min), followed bylyophilization to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-(4-methylpiperazin-1-yl)pyrimidin-4-amine(49.11 mg, 113.81 μmol, 70.5% yield, 100.0% purity) as a white solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 10.82 (br s, 1H), 9.30 (s, 1H), 8.64 (d,J=2.9 Hz, 1H), 8.32 (d, J=10.1 Hz, 1H), 7.59 (d, J=7.7 Hz, 1H), 7.34 (d,J=7.9 Hz, 1H), 7.20 (d, J=2.0 Hz, 1H), 7.07 (t, J=7.2 Hz, 1H), 6.98 (t,J=7.4 Hz, 2H), 5.59 (br s, 1H), 3.60 (br s, 2H), 3.51 (br s, 4H), 2.97(t, J=7.3 Hz, 2H), 2.37 (d, J=4.2 Hz, 4H), 2.20 (s, 3H); ES-LCMS m/z432.2 [M+H]⁺.

Example 67

Synthesis of I-68

Synthetic Scheme:

Step 1:[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]-(4-methylpiperazin-1-yl)methanone(I-68)

To a solution of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (50 mg, 120.07 μmol, 1 eq) and 1-methylpiperazine (18.04 mg, 180.11μmol, 19.98 μL, 1.5 eq) in DCM (5 mL) was added HATU (54.79 mg, 144.08μmol, 1.2 eq) and DIEA (46.55 mg, 360.21 μmol, 62.74 μL, 3.0 eq). Themixture was stirred at 25° C. for 16 h. LC-MS showed 42% of startingmaterial was remained and 51% of desired compound was detected.1-methylpiperazine (18.04 mg, 180.11 μmol, 19.98 μL, 1.5 eq), HATU(54.79 mg, 144.08 μmol, 1.2 eq), DIEA (46.55 mg, 360.21 μmol, 62.74 μL,3.0 eq) and DMF (2 mL) was added. The mixture was stirred at 25° C. for5 h. The reaction mixture was diluted with H₂O (20 mL), extracted withDCM (20 mL×3). The combined organic layers were dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue which purified by preparative HPLC (column: Phenomenex Gemini150*25 mm*10 um; mobile phase: [water (0.05% ammonia hydroxidev/v)-ACN]; B %: 36%-66%, 10 min), followed by lyophilization to yield[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]-(4-methylpiperazin-1-yl)methanone (15.94 mg, 31.97 μmol,26.6% yield, 100.0% purity) as a light yellow solid. ¹H NMR (400 MHz,CD₃OD) δ ppm 8.63 (s, 1H), 8.48 (d, J=2.9 Hz, 1H), 8.26 (s, 1H),7.71-7.62 (m, 2H), 7.19-7.13 (m, 1H), 7.07-6.95 (m, 3H), 6.05 (s, 1H),3.90 (t, J=6.2 Hz, 2H), 3.75 (br s, 4H), 3.19 (t, J=6.1 Hz, 2H), 2.54(br s, 4H), 2.33 (s, 3H); ES-LCMS m/z 499.2 [M+H]⁺.

Example 68

Synthesis of I-69

Synthetic Scheme:

Step 1:N-[2-(5-Fluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-amine(I-69)

To a solution of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine (50mg, 164.55 μmol, eq) in i-PrOH (3 mL) was added DIEA (106.33 mg, 822.73μmol, 143.31 μL, 5 eq) and 2-(5-fluoro-2-methyl-1H-indol-3-yl)ethanamine(37.96 mg, 197.46 μmol, 1.2 eq). The mixture was stirred at 60° C. for16 h. The reaction mixture was concentrated under reduced pressure togive a residue which was purified by preparative HPLC (column:Phenomenex Synergi C18 150*25 mm*10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 45%-75%, 10 min) followed by lyophilization to yieldN-[2-(5-fluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-amine(32.52 mg, 58.40 μmol, 35.5% yield, 100.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.35 (s, 1H), 8.93 (s, 1H), 8.77(s, 1H), 8.62 (d, J=9.3 Hz, 1H), 7.15-6.94 (m, 2H), 6.63 (t, J=8.0 Hz,1H), 5.10-4.99 (m, 1H), 4.04 (m, 2H), 3.09 (t, J=6.6 Hz, 2H), 2.30 (s,3H), 1.71 (d, J=6.8 Hz, 6H); ES-LCMS m/z 448.0 [M+H]⁺.

Example 69

Synthesis of I-70

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-70)

To a solution of 2-(1H-indol-3-yl)ethanamine (29.28 mg, 182.74 μmol, 1.2eq) in i-PrOH (3 mL) was added DIEA (59.04 mg, 456.85 μmol, 79.57 μL, 3eq) and7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (40mg, 152.28 μmol, 1 eq). The mixture was stirred at 50° C. for 16 h underN₂ atmosphere. The reaction mixture was concentrated under reducedpressure to remove i-PrOH to give a residue which was purified bypreparative HPLC (column: Phenomenex Synergi C18 150*25 mm*10 um; mobilephase: [water(0.05% HCl)-ACN]; B %: 35%-65%, 10 min) followed bylyophilization to yield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(32.43 mg, 65.15 μmol, 42.8% yield, 99.6% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.72 (d, J=2.6 Hz, 1H), 8.35 (s,1H), 8.10 (s, 1H), 7.56-7.52 (m, 1H), 7.50-7.45 (m, 1H), 7.16-7.10 (m,1H), 6.98 (s, 1H), 6.97-6.92 (m, 1H), 6.85 (m, 1H), 5.76 (s, 1H),4.06-3.99 (m, 2H), 3.24-3.19 (m, 2H), 2.30 (s, 3H); ES-LCMS m/z 387.0[M+H]⁺.

Example 70

Synthesis of I-71

Synthetic Scheme:

Step 1:N-[2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-amine(I-71)

To a solution of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine(41.67 mg, 137.12 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (53.16 mg,411.37 μmol, 71.65 μL, 3.0 eq),2-(7-fluoro-2-methyl-1H-indol-3-yl)ethanamine (37.66 mg, 164.55 μmol,1.2 eq). The mixture was stirred at 60° C. for 12 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by preparative HPLC (column: Phenomenex Gemini 150×25 mm×10μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 46%-76%, 10 min) followedby lyophilization to yieldN-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-amine(27.83 mg, 49.98 μmol, 36.4% yield, 100.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.30 (s, 1H), 8.80 (s, 1H), 8.74(dd, J=1.1, 2.9 Hz, 1H), 8.61-8.56 (m, 1H), 7.28 (d, J=7.9 Hz, 1H), 6.84(dt, J=4.7, 7.9 Hz, 1H), 6.63 (dd, J=7.9, 11.7 Hz, 1H), 5.03 (td, J=6.9,13.6 Hz, 1H), 4.04 (m, 2H), 3.12 (t, J=6.6 Hz, 2H), 2.30 (s, 3H), 1.70(d, J=6.8 Hz, 6H); ES-LCMS m/z 447.9 [M+H]⁺.

Example 71

Synthesis of I-72

Synthetic Scheme:

Step 1: 2,4-Dichloro-6-isopropoxy-pyrimidine

To a solution of i-PrOH (344.04 mg, 5.72 mmol, 438.27 μL, 1.05 eq) inTHE (8 mL) was added NaH (261.69 mg, 6.54 mmol, 60% in mineral oil, 1.2eq). The mixture was stirred at 0° C. for 30 min.2,4,6-Trichloropyrimidine (1 g, 5.45 mmol, 625.00 μL, 1.0 eq) was addedinto the above solution and the mixture was stirred at 15° C. for 12 h.The reaction mixture was concentrated under reduced pressure to removeTHF. The residue was diluted with water (150 mL), extracted with EtOAc(50 mL×3). The combined organic layers were dried over Na₂SO₄, filteredand the filtrate was concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=100/1 to 50/1, TLC: PE/EtOAc=100/1, R_(f)=0.20) to yieldmixture 4,6-dichloro-2-isopropoxy-pyrimidine (347 mg, crude) and2,4-dichloro-6-isopropoxy-pyrimidine (173 mg, crude) as colorless oil.¹H NMR (400 MHz, CDCl₃) δ ppm 7.00 (s, 1H), 5.35-5.23 (m, 1H), 1.40 (d,J=6.2 Hz, 6H); ES-LCMS m/z 207.1, 209.1 [M+H]⁺.

Step 2:4-Chloro-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-2-amine

To a solution of (2-(1H-indol-3-yl)ethanamine (680.92 mg, 4.25 mmol, 2.2eq) and 2,4-dichloro-6-isopropoxy-pyrimidine (100 mg, 482.96 μmol, 0.25eq) in i-PrOH (5 mL) was added DIEA (749.01 mg, 5.80 mmol, 1.01 mL, 3.0eq) and 4,6-dichloro-2-isopropoxy-pyrimidine (200 mg, 965.92 μmol, 0.5eq). The mixture was stirred at 26° C. for 12 h. The reaction mixturewas concentrated under reduced pressure to give the residue which waspurified by flash silica gel chromatography (from PE/EtOAc=100/1 to 3/1,TLC: PE/EtOAc=3/1, R_(f)=0.55, R_(f2)=0.45) to yield4-chloro-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-2-amine (90mg, 220.37 μmol, 11.4% yield, 81.0% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.05 (s, 1H), 7.68-7.61 (m, 1H), 7.39 (d, J=8.2Hz, 1H), 7.25-7.19 (m, 1H), 7.16-7.10 (m, 1H), 7.05 (s, 1H), 5.97 (s,1H), 3.78-3.69 (m, 2H), 3.10-3.03 (m, 2H), 1.33 (d, J=5.3 Hz, 6H);ES-LCMS m/z 331.1, 332.1 [M+H]⁺.

Step 3:4-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-2-amine(I-72)

4-Chloro-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-2-amine(90.00 mg, 220.37 μmol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (62.10mg, 440.73 μmol, 2 eq), Cs₂CO₃ (215.40 mg, 661.10 μmol, 3.0 eq) andPd(dppf)Cl₂ (16.12 mg, 22.04 μmol, 0.1 eq) were taken up into amicrowave tube in 1,4-dioxane (3 mL) and water (0.6 mL). The sealed tubewas heated at 120° C. for 30 min under microwave. The reaction mixturewas diluted with EtOAc (20 mL) and filtered through a pad of celite. Thefiltrate was concentrated under reduced pressure to give a residue whichwas purified by prep-HPLC (HCl condition, column: column: PhenomenexSynergi C18 150×30 mm×4 um; mobile phase: [water (0.05% HCl)-ACN]; B %:37%-67%, 12 min) and the desired fraction was lyophilized to yield4-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-2-amine(52.48 mg, 104.79 μmol, 47.5% yield, 100.0% purity, 3HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.75 (m, 2H), 8.06 (s, 1H),7.60 (d, J=7.8 Hz, 1H), 7.33 (d, J=8.0 Hz, 1H), 7.14 (s, 1H), 7.12-7.06(m, 1H), 7.04-6.99 (m, 1H), 6.62 (s, 1H), 5.18 (s, 1H), 3.92 (t, J=6.5Hz, 2H), 3.15 (t, J=6.5 Hz, 2H), 1.36 (d, J=6.3 Hz, 6H); ES-LCMS m/z392.2 [M+H]⁺.

Example 72

Synthesis of I-73

Synthetic Scheme:

Step 1: 2-Methyl-3-oxo-propanenitrile

To a solution of DIPA (1.84 g, 18.16 mmol, 2.57 mL, 1 eq) in THE (20 mL)was added n-BuLi (2.5 M in n-hexane, 7.63 mL, 1.05 eq). The mixture wasstirred at −65° C. for 30 min. The solution of propanenitrile (1 g,18.16 mmol, 1.30 mL, 1 eq) in THE (10 mL) was added into the abovemixture dropwise. The mixture was stirred at −65° C. for 30 min under N₂atmosphere. A solution of ethyl formate (1.41 g, 19.06 mmol, 1.53 mL,1.05 eq) in THF (10 mL) was added dropwise and it was stirred at −65° C.for 3 h. TLC (PE/EtOAc=1/1, R_(f)=0.45) showed one major new spot wasdetected. The reaction mixture was quenched by addition 1 N HCl solution(50 mL) at −65° C., extracted with EtOAc (50 mL×3). The combined organiclayers were dried over Na₂SO₄, filtered and the filtrate wasconcentrated under reduced pressure to give2-methyl-3-oxo-propanenitrile (1.1 g, crude) was obtained as yellow oilwhich was used in the next step without further purification. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.60 (s, 1H), 3.54 (d, J=7.5 Hz, 1H), 1.55 (s,3H).

Step 2: 4-Methyl-1H-pyrazol-5-amine

To a solution of 2-methyl-3-oxo-propanenitrile (1.1 g, 13.24 mmol, 1 eq)in EtOH (12 mL) was added AcOH (1.39 g, 23.17 mmol, 1.32 mL, 1.75 eq)and hydrazine (550.00 mg, 17.16 mmol, 620.77 μL, 1.3 eq). The mixturewas stirred at 90° C. for 12 h. TLC (PE/EtOAc=1/1, R_(f)=0.10) showedone major new spot was detected. The reaction mixture was concentratedunder reduced pressure to give a residue which was diluted with NaHCO₃solution (100 mL), extracted with EtOAc (50 mL×3). The combined organiclayers were dried over Na₂SO₄, filtered and the filtrate wasconcentrated under reduced pressure to give a residue which was purifiedby flash silica gel chromatography (from DCM/MeOH=100/1 to 10/1, TLC:PE/EtOAc=10/1, R_(f)=0.75) to yield compound 4-methyl-1H-pyrazol-5-amine(230 mg, 2.37 mmol, 17.9% yield, crude) as yellow oil. ¹H NMR (400 MHz,CDCl₃) δ ppm 7.13 (s, 1H), 1.94-1.92 (m, 3H).

Step 3: 5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-ol

To a solution of methyl 3-(5-fluoro-3-pyridyl)-3-oxo-propanoate (400 mg,1.99 mmol, 1 eq) in AcOH (6 mL) was added 4-methyl-1H-pyrazol-5-amine(230 mg, 2.37 mmol, 1.19 eq). The mixture was stirred at 120° C. for 2h. The reaction mixture was concentrated under reduced pressure to give5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-ol (500 mg,crude) was obtained as yellow oil which was used in the next stepwithout further purification. ES-LCMS m/z 245.2 [M+H]⁺.

Step 4:7-Chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine

To a solution of5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-ol (500 mg,655.14 μmol, 1 eq) in POCl₃ (5 mL) was stirred at 120° C. for 2 h. Thereaction mixture was concentrated under reduced pressure to removePOCl₃. The residue was purified by flash silica gel chromatography (fromPE/EtOAc=100/1 to 2/1, TLC: PE/EtOAc=3/1, R_(f)=0.40) to yield7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (145mg, 552.02 μmol, 84.3% yield, 100.0% purity) as yellow solid ¹H NMR (400MHz, CDCl₃) δ ppm 9.10 (s, 1H), 8.60 (d, J=2.6 Hz, 1H), 8.24 (dd, J=1.8,9.3 Hz, 1H), 8.14 (s, 1H), 7.39 (s, 1H), 2.47 (s, 3H); ES-LCMS m/z262.9, 264.9 [M+H]⁺.

Step 5:5-(5-Fluoro-3-pyridyl)-3-methyl-N-[2-(2,5,7-trifluoro-1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-73)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (40mg, 152.28 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (157.45 mg, 1.22mmol, 212.19 μL, 8.0 eq) and2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethanamine (54.87 mg, 182.74μmol, 1.2 eq, oxalic acid salt). The mixture was stirred at 50° C. for12 h. The reaction mixture was concentrated under reduced pressure togive the residue which was purified by preparative HPLC (HCl condition;column: Phenomenex Gemini 150×25 mm×10 um; mobile phase: [water(0.05%HCl)-ACN]; B %: 42%-62%, 10 min) and the desired fraction waslyophilized to yield compound5-(5-fluoro-3-pyridyl)-3-methyl-N-[2-(2,5,7-trifluoro-1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(15.56 mg, 27.91 μmol, 18.3% yield, 98.6% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.75 (d, J=2.4 Hz, 1H), 8.45 (s,1H), 8.11 (s, 1H), 7.66-7.61 (m, 1H), 7.00 (dd, J=2.2, 9.3 Hz, 1H), 6.44(ddd, J=2.2, 9.5, 11.2 Hz, 1H), 5.75 (s, 1H), 4.00 (t, J=5.7 Hz, 2H),3.16-3.10 (m, 2H), 2.31 (s, 3H), 2.15 (s, 3H); ES-LCMS m/z 437.2 [M+H]⁺.

Example 73

Synthesis of I-74

Synthetic Scheme:

Step 1:N-[2-(5,7-Difluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidin-4-amine(I-74)

4-Chloro-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidine (90 mg, 322.77μmol, 1.0 eq), 2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethanamine (145.37mg, 484.15 μmol, 1.5 eq, oxalic acid salt) and DIEA (333.71 mg, 2.58mmol, 449.75 uL, 8 eq) in i-PrOH (5 mL) were taken up into a microwavetube. The sealed tube was heated at 150° C. for 6 h under microwave. Thereaction mixture was concentrated under reduced pressure to give theresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=100/1 to 2/1, TLC: PE/EtOAc=3/1, R_(f)=0.50) and thenre-purified by preparative HPLC (HCl condition; column: PhenomenexGemini 150×25 mm×10 um; mobile phase: [water (0.05% HCl)-ACN]; B %:60%-90%, 10 min) and the desired fraction was lyophilized to yieldN-[2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidin-4-amine(26.48 mg, 48.07 μmol, 14.9% yield, 100% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.10 (s, 1H), 8.71 (d, J=2.6 Hz,1H), 8.27 (s, 1H), 6.99 (d, J=9.3 Hz, 1H), 6.54 (t, J=9.9 Hz, 1H), 5.69(s, 1H), 5.05 (s, 1H), 3.74 (m, 2H), 2.97 (t, J=6.6 Hz, 2H), 2.34 (s,3H), 1.37 (d, J=6.2 Hz, 6H); ES-LCMS m/z 442.2 [M+H]⁺.

Example 74

Synthesis of I-75a, I-75b and I-75c

Step 1: tert-Butyl N-(2,3,4,9-tetrahydro-1H-carbazol-3-yl)carbamate

To a solution of tert-butyl N-(4-oxocyclohexyl)carbamate (1 g, 4.69mmol, 1.00 mL, 1 eq) in DCM (30 mL) was added MgSO₄ (564.38 mg, 4.69mmol, 1 eq) and phenylhydrazine (507.05 mg, 4.69 mmol, 460.96 μL, 1 eq).The mixture was stirred at 28° C. for 2 h. Then MgSO₄ was filtered off,the filtrate was evaporated under reduced pressure. The resulting brownoil was dissolved in toluene (20 mL), ZnCl₂ (3.20 g, 23.44 mmol, 5 eq)was added, the mixture was heated at 110° C. for 4 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by flash silica gel chromatography (from PE/EtOAc=100/1 to3/1, TLC: PE/EtOAc=5/1, R_(f)=0.30) to yield compound tert-butylN-(2,3,4,9-tetrahydro-1H-carbazol-3-yl)carbamate (700 mg, 1.96 mmol,41.7% yield, 80.0% purity) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δppm 7.33 (d, J=7.5 Hz, 1H), 7.23 (td, J=0.9, 7.9 Hz, 1H), 7.04-6.97 (m,1H), 6.97-6.90 (m, 1H), 3.92-3.79 (m, 1H), 2.92-2.78 (m, 2H), 2.57-2.31(m, 2H), 1.93-1.68 (m, 2H), 1.47 (s, 9H); ES-LCMS m/z 231.1 [M-t-Bu+H]⁺.

Step 2: 2,3,4,9-Tetrahydro-1H-carbazol-3-amine

To a solution of tert-butylN-(2,3,4,9-tetrahydro-1H-carbazol-3-yl)carbamate (700 mg, 1.96 mmol, 1eq) in DCM (10 mL) was added TFA (3.08 g, 27.01 mmol, 2.0 mL, 13.81 eq).The mixture was stirred at 28° C. for 2 h. TLC (PE/EtOAc=5/1, R_(f)=0)indicated the starting material was consumed completely and one new spotformed. The reaction mixture was concentrated under reduced pressure togive the residue which was diluted with water (30 mL), adjusted pH=10 by1% NaOH solution. The mixture was extracted with EtOAc (30 mL×3). Thecombined organic layers were dried over Na₂SO₄, filtered and thefiltrate was concentrated under reduced pressure to give2,3,4,9-tetrahydro-1H-carbazol-3-amine (400 mg, 1.80 mmol, 92.3% yield,84.0% purity) was obtained as a black brown solid which was used in thenext step without further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm7.80 (s, 1H), 7.44 (d, J=7.7 Hz, 1H), 7.27-7.23 (m, 1H), 7.13-7.04 (m,2H), 3.34-3.24 (m, 1H), 3.07-2.97 (m, 1H), 2.81 (t, J=6.4 Hz, 2H),2.50-2.43 (m, 1H), 2.13-2.05 (m, 1H), 1.84-1.76 (m, 1H); ES-LCMS m/z187.0 [M+H]⁺.

Step 3:(3S)—N-[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-75b) &(3R)—N-[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-75a)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(153.06 mg, 515.96 μmol, 1.0 eq) in i-PrOH (10 mL) was added DIEA(200.05 mg, 1.55 mmol, 269.60 μL, 3.0 eq) and2,3,4,9-tetrahydro-1H-carbazol-3-amine (145.09 mg, 654.35 μmol, 1.27eq). The mixture was stirred at 50° C. for 15 h. The reaction mixturewas concentrated under reduced pressure to give a residue which waspurified by flash silica gel chromatography (from PE/EtOAc=100/1 to 2/1,TLC: PE/EtOAc=3/1, R_(f)=0.80). The compounds were separated by SFC(condition: column: AD (250 mm×30 mm, 5 um); mobile phase: [0.1% NH₃H₂OEtOH]; B %: 45%-45%, min). The solution after separation wereconcentrated to afford the crude products which were purified bypreparative HPLC (HCl condition; column: Phenomenex Synergi C18 150×30mm×4 um; mobile phase: [water(0.05% HCl)-ACN]; B %: 60%-90%, 12 min),followed by lyophilization to yield an enantiomer (36.18 mg, 65.79 μmol,12.8% yield, 100.0% purity, 3HCl salt) (Rt=4.768 min, ee %=100.0 and[α]²⁹ _(D)=−3.430 (i-ProH, c=0.107 g/100 mL)) as a yellow solid; ¹H NMR(400 MHz, CD₃OD) δ ppm 9.00 (s, 1H), 8.78 (s, 1H), 8.36-8.29 (m, 1H),8.24 (d, J=1.3 Hz, 1H), 7.38 (d, J=7.7 Hz, 1H), 7.27 (d, J=7.9 Hz, 1H),7.08-6.93 (m, 3H), 4.63 (s, 1H), 3.43-3.32 (m, 2H), 3.18-2.93 (m, 3H),2.43-2.26 (m, 2H), 1.40 (d, J=6.8 Hz, 6H); ES-LCMS m/z 441.2 [M+H]⁺; andthe other enantiomer (36.49 mg, 66.36 μmol, 12.9% yield, 100.0% purity,3HCl salt) (Rt=5.778 min, ee %=96.4 and [α]²⁹ _(D)=+3.121 (i-ProH,c=0.104 g/100 mL)) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.03(s, 1H), 8.73 (d, J=2.6 Hz, 1H), 8.36-8.30 (m, 1H), 8.18 (s, 1H), 7.38(d, J=7.7 Hz, 1H), 7.29-7.25 (m, 1H), 7.04 (dt, J=1.2, 7.6 Hz, 1H),7.00-6.93 (m, 2H), 4.60 (s, 1H), 3.40-3.32 (m, 2H), 3.16-2.92 (m, 3H),2.43-2.24 (m, 2H), 1.40 (d, J=6.8 Hz, 6H); ES-LCMS m/z 441.3 [M+H]⁺.

Example 75

Synthesis of I-77

Synthetic Scheme:

Step 1: 5-Fluoro-2-methyl-1H-indole-3-carbaldehyde

To DMF (40 mL) was added POCl₃ (4.11 g, 26.82 mmol, 2.49 mL, 2 eq)dropwise at −20° C. over a period of 10 min under N₂ atmosphere. Afterbeing stirred for 1 h, 5-fluoro-2-methyl-1H-indole (2 g, 13.41 mmol, 1eq) was added to the above solution during which the temperature wasmaintained below −20° C. The reaction mixture was warmed to 15° C. andstirred for 1 h. The reaction mixture was quenched by saturated aqueousNaHCO₃ (100 mL) and extracted with EtOAc (50 mL×3). The combine organiclayers were dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by flash silicagel chromatography (from PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=1/1,R_(f)=0.67) to yield 5-fluoro-2-methyl-1H-indole-3-carbaldehyde (1.3 g,5.73 mmol, 42.7% yield, 78.1% purity) as a yellow solid. ¹H NMR (400MHz, CDCl₃) δ ppm 10.17 (s, 1H), 8.39 (br s, 1H), 7.93 (d, J=9.0 Hz,1H), 7.08-6.89 (m, 1H), 2.75 (s, 3H); ES-LCMS m/z 178.0 [M+H]⁺.

Step 2: 5-Fluoro-2-methyl-3-[(E)-2-nitrovinyl]-1H-indole

To a solution of 5-fluoro-2-methyl-1H-indole-3-carbaldehyde (1.1 g, 4.85mmol, 1 eq) in nitronethane (45 mL) was added NH₄OAc (1.12 g, 14.54mmol, 3 eq). The mixture was stirred at 110° C. for 16 h. The reactionmixture was concentrated under reduced pressure to remove nitronethane.The residue was extracted with EtOAc (50 mL×3). The combine organiclayers were dried over Na₂SO₄, filtered and concentrated under reducedpressure to give 5-fluoro-2-methyl-3-[(E)-2-nitrovinyl]-1H-indole (0.92g, crude) as a brown solid which was used in the next step withoutfurther purification; ES-LCMS m/z 221.0 [M+H]⁺.

Step 3: 2-(5-Fluoro-2-methyl-1H-indol-3-yl)ethanamine

To a solution of 5-fluoro-2-methyl-3-[(E)-2-nitrovinyl]-1H-indole (1.40g, 6.36 mmol, 1 eq) in THF (30 mL) was added LAH (1 M, 31.79 mL, 5 eq)at 0° C. After addition, the mixture was stirred at 80° C. for 2 h. Thereaction mixture was diluted with THE (10 mL) and then was quenched byaddition water (1.4 mL), 10% NaOH (1.4 mL) and water (4.2 mL) insequence at 0° C. After being stirred for 30 min, the mixture wasfiltered through celite. The filtrate was concentrated under reducedpressure to give 2-(5-fluoro-2-methyl-1H-indol-3-yl)ethanamine (1.0 g,crude) as yellow oil which was used in the next step directly withoutfurther purification. ¹H NMR (400 MHz, CD₃OD) δ ppm 7.14 (dd, J=8.8 Hz,1H), 7.07 (dd, J=10.0 Hz, 1H), 6.72 (td, J=5.0, 9.1 Hz, 1H), 2.85-2.74(m, 4H), 2.33 (s, 3H); ES-LCMS m/z 192.1 [M+H]⁺.

Step 4:N-[2-(5-Fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-77)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(64.30 mg, 216.75 μmol, 1 eq) in i-PrOH (5 mL) was added DIEA (140.07mg, 1.08 mmol, 188.77 μL, 5 eq) and2-(5-fluoro-2-methyl-1H-indol-3-yl)ethanamine (50 mg, 260.10 μmol, 1.2eq) The mixture was stirred at 60° C. for 16 h. The reaction mixture wasconcentrated under reduced pressure to remove i-PrOH to give a residuewhich was purified by preparative HPLC (Column: Phenomenex Gemini 150*25mm*10 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 50%-80%, 7 min).followed by lyophilization to yieldN-[2-(5-fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(17.49 mg, 31.46 umol, 14.5% yield, 100.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.75 (d, J=2.4 Hz, 1H), 8.40 (s,1H), 8.21 (s, 1H), 7.54 (d, J=8.6 Hz, 1H), 7.07 (dd, J=2.3, 9.8 Hz, 1H),6.95 (dd, J=4.4, 8.6 Hz, 1H), 6.59 (J=2.4, 9.0 Hz, 1H), 5.71 (s, 1H),4.03-3.95 (m, 2H), 3.28-3.20 (m, 1H), 3.16-3.09 (m, 2H), 2.16 (s, 3H),1.34 (d, J=6.8 Hz, 6H); ES-LCMS m/z 447.1 [M+H]⁺.

Example 76

Synthesis of I-78

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxamide(I-78)

A mixture of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (35 mg, 84.05 μmol, 1 eq), HATU (63.92 mg, 168.10 μmol, 2 eq), DIEA(32.59 mg, 252.16 μmol, 43.92 μL, 3 eq) and NH₄Cl (13.49 mg, 252.16μmol, 3 eq) in DMF (5 mL) was stirred at 25° C. for 3 h. The mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um; mobilephase: [water(0.05% HCl)-ACN]; B %: 36%-56%, 10 min), lyophilized togive5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxamide(16.47 mg, 31.08 μmol, 37.0% yield, 99.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.85 (s, 1H), 8.73 (s, 1H), 8.51(s, 1H), 8.03 (d, J=9.2 Hz, 1H), 7.72-7.69 (m, 1H), 7.12 (d, J=8.4 Hz,1H), 7.04-7.01 (m, 2H), 6.95 (s, 1H), 6.08 (s, 1H), 3.98 (t, J=5.6 Hz,2H), 3.22 (t, J=5.6 Hz, 2H); ES-LCMS m/z 416.2 [M+H]⁺.

Example 77

Synthesis of I-79

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-9-isopropyl-N-[2-(2-methyl-1H-indol-3-yl)ethyl]purin-6-amine(I-79)

A mixture of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine (52.08mg, 171.40 μmol, 1 eq), 2-(2-methyl-1H-indol-3-yl)ethanamine (29.87 mg,171.40 μmol, 1 eq) and DIEA (66.46 mg, 514.21 μmol, 89.57 μL, 3 eq) ini-PrOH (3 mL) was stirred at 55° C. for 19 h. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Synergi C18 150*30 mm*4 um;mobile phase: [water (0.05% HCl)-ACN]; B %: 42%-72%, 12 min) followed bylyophilization to yield2-(5-fluoro-3-pyridyl)-9-isopropyl-N-[2-(2-methyl-1H-indol-3-yl)ethyl]purin-6-amine(18.99 mg, 34.91 μmol, 20.4% yield, 99.1% purity, 3 HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.27-9.11 (m, 2H), 8.85 (s,1H), 8.68 (d, J=8.78 Hz, 1H), 7.51-7.43 (m, 1H), 7.07-7.00 (m, 1H),6.92-6.83 (m, 2H), 5.13-5.00 (m, 1H), 4.07 (s, 2H), 3.11 (t, J=6.53 Hz,2H), 2.27 (s, 3H), 1.73 (d, J=6.53 Hz, 6H); ES-LCMS m/z 430.2 [M+H]⁺.

Example 78

Synthesis of I-80

Synthetic Scheme:

Step 1: 4,6-Dichloro-2-isopropoxy-pyrimidine

To a suspension of NaH (248.00 mg, 6.20 mmol, 60% in mineral oil, 1.41eq) in dry THE (10 mL) was added i-PrOH (277.91 mg, 4.62 mmol, 354.03μL, 1.05 eq) under ice bath and N₂ atmosphere. After being stirred for30 min, the suspension was cooled to −60° C. and4,6-dichloro-2-methylsulfonyl-pyrimidine (1 g, 4.40 mmol, 625.00 μL, 1.0eq) in dry THE (10 mL) was added dropwise and kept the temperature below−55° C. The resulting mixture was stirred for 1 h at −55° C. Thereaction mixture was poured into water (100 mL) slowly and extractedwith EtOAc (30 mL×3). The combined organic layers were washed withbrine, dried over Na₂SO₄ and concentrated to yield4,6-dichloro-2-isopropoxy-pyrimidine (565 mg, 2.56 mmol, 58.2% yield,94% purity) as yellow oil which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.02-6.96 (m, 1H),5.29 (m, 1H), 1.40 (d, J=6.0 Hz, 6H); ES-LCMS m/z 206.9, 208.9 [M+H]⁺.

Step 2:6-Chloro-N-[2-(1H-indol-3-yl)ethyl]-2-isopropoxy-pyrimidin-4-amine

To a solution of 4,6-dichloro-2-isopropoxy-pyrimidine (200 mg, 907.96μmol, 0.5 eq) in i-PrOH (6 mL) was added DIEA (704.07 mg, 5.45 mmol,948.88 μL, 3.0 eq) and 2-(1H-indol-3-yl)ethanamine (329.00 mg, 2.05mmol, 1.13 eq). The mixture was stirred at 26° C. for 12 h. The reactionmixture was concentrated under reduced pressure to give the residuewhich was purified by flash silica gel chromatography (fromPE/EtOAc=100/1 to 3/1, TLC: PE/EtOAc=3/1, R_(f)=0.45) to yield6-chloro-N-[2-(1H-indol-3-yl)ethyl]-2-isopropoxy-pyrimidin-4-amine (290mg, 841.57 μmol, 46.3% yield, 96% purity) as yellow oil. ¹H NMR (400MHz, CDCl₃) δ ppm 8.10 (s, 1H), 7.60 (d, J=7.7 Hz, 1H), 7.43-7.37 (m,1H), 7.24 (dt, J=1.0, 7.6 Hz, 1H), 7.18-7.12 (m, 1H), 7.05 (d, J=2.4 Hz,1H), 5.95 (s, 1H), 5.26-5.17 (m, 1H), 3.68 (s, 2H), 3.08 (t, J=6.6 Hz,2H), 1.35 (d, J=6.2 Hz, 6H); ES-LCMS m/z 330.9, 332.0 [M+H]⁺.

Step 3:6-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-2-isopropoxy-pyrimidin-4-amine(I-80)

6-Chloro-N-[2-(1H-indol-3-yl)ethyl]-2-isopropoxy-pyrimidin-4-amine (90mg, 261.18 μmol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (92.00 mg,652.94 μmol, 2.5 eq), Cs₂CO₃ (255.29 mg, 783.53 μmol, 3.0 eq) andPd(dppf)Cl₂ (19.11 mg, 26.12 μmol, 0.1 eq) were taken up into amicrowave tube in water (1.2 mL) and 1,4-dioxane (6 mL). The sealed tubewas heated at 120° C. for 30 min under microwave. The reaction mixturewas diluted with EtOAc (15 mL) and filtered through a pad of celite. Thefiltrate was concentrated under reduced pressure to give the residuewhich was purified by preparative HPLC (HCl condition, column: column:Phenomenex Gemini 150×25 mm×10 um; mobile phase: [water(0.05% HCl)-ACN];B %: 25%-55%, 10 min) and the desired fraction was lyophilized to yield6-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-2-isopropoxy-pyrimidin-4-amine(43.81 mg, 87.48 μmol, 33.5% yield, 100% purity, 3 HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD+Na₂CO₃) δ ppm 8.92 (s, 1H), 8.53 (m, 1H),8.12 (m, 1H), 7.61 (m, 1H), 7.33 (m, 1H), 7.09 (d, J=7.0 Hz, 2H), 7.02(m, 1H), 6.57 (s, 1H), 5.38-5.26 (m, 1H), 3.77 (m, 2H), 3.09 (t, J=7.0Hz, 2H), 1.40 (d, J=5.8 Hz, 6H); ES-LCMS m/z 392.0 [M+H]⁺.

Example 79

Synthesis of I-81

Synthetic Scheme:

Step 1: Methyl 3-(5-bromo-3-pyridyl)-3-oxo-propanoate

A mixture of 5-bromopyridine-3-carboxylic acid (1 g, 4.95 mmol, 1 eq)and CDI (1.20 g, 7.43 mmol, 1.5 eq) in THE (10 mL) was stirred at 20° C.for 1 h, TEA (500.93 mg, 4.95 mmol, 689.03 μL, 1 eq) was added, themixture was stirred at 20° C. for 1 h, then(3-methoxy-3-oxo-propanoyl)oxypotassium(1+) (1.55 g, 9.90 mmol, 2 eq)and MgCl₂ (942.66 mg, 9.90 mmol, 406.32 μL, 2 eq) were added. Themixture was stirred at 20° C. for 11 h. The reaction mixture was adjustpH to 2 with 2N HCl, extracted with EtOAc (20 mL×3). The combinedorganic layers were washed with brine (20 mL), dried over Na₂SO₄,filtered and concentrated under reduced pressure to give a residue whichwas purified by flash silica gel chromatography (from PE/EtOAc=10/1 to5/1, TLC: PE/EtOAc=5/1, R_(f)=0.58) to yield methyl3-(5-bromo-3-pyridyl)-3-oxo-propanoate (554 mg, 1.50 mmol, 30.4% yield,70% purity) as a white solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.07-9.06(m, 1H), 8.95 (d, J=1.6 Hz, 1H), 8.89 (d, J=2.0 Hz, 1H), 8.76 (d, J=2.4Hz, 1H), 8.53-8.52 (m, 1H), 8.42 (t, J=2.0 Hz, 1H), 5.97 (s, 1H), 3.82(s, 3H), 3.74 (s, 3H); ES-LCMS m/z 259.9, 261.9 [M+H]⁺.

Step 2: 5-(5-Bromo-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-ol

A mixture of methyl 3-(5-bromo-3-pyridyl)-3-oxo-propanoate (300 mg,813.74 μmol, 1 eq) and 4-isopropyl-1H-pyrazol-5-amine (122.23 mg, 976.49μmol, 1.2 eq) in AcOH (10 mL) was stirred at 120° C. for 0.5 h. Themixture was concentrated under reduced pressure to give5-(5-bromo-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-ol (450 mg,crude) as brown oil which was used in next step without furtherpurification. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.99-8.88 (m, 1H), 8.74-8.73(m, 1H), 8.39-8.37 (m, 2H), 7.79 (s, 1H), 3.21-3.20 (m, 1H), 1.39-1.33(m, 6H); ES-LCMS m/z 333.0, 335.0 [M+H]⁺.

Step 3:5-(5-Bromo-3-pyridyl)-7-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine

A solution of5-(5-bromo-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-ol (450 mg,1.35 mmol, 1 eq) in POCl₃ (8.2 g, 53.48 mmol, 4.97 mL, 39.60 eq) wasstirred at 110° C. for 1 h. The mixture was concentrated under reducedpressure to give a residue which was purified by flash silica gelchromatography (from PE/EtOAc=10/1 to 5/1, TLC: PE/EtOAc=5/1,R_(f)=0.58) to yield5-(5-bromo-3-pyridyl)-7-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine(331 mg, 847.20 μmol, 62.7% yield, 90% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.20 (s, 1H), 8.79 (d, J=2.4 Hz, 1H), 8.59 (t,J=2.4 Hz, 1H), 8.14 (s, 1H), 7.37 (s, 1H), 3.42 (s, 1H), 1.45 (d, J=6.8Hz, 6H); ES-LCMS m/z 350.9, 352.9 [M+H]⁺.

Step 4:5-(5-Bromo-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine

A mixture of5-(5-bromo-3-pyridyl)-7-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine (60mg, 153.57 μmol, 1 eq), 2-(1H-indol-3-yl)ethanamine (36.91 mg, 230.36μmol, 1.5 eq) and DIEA (59.54 mg, 460.71 μmol, 80.25 μL, 3 eq) in i-PrOH(10 mL) was stirred at 80° C. for 2 h. The mixture was concentratedunder reduced pressure to give a residue which was purified bypreparative TLC (SiO₂, PE/EtOAc=1/1, R_(f)=0.28) to yield5-(5-bromo-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(70 mg, 117.80 μmol, 76.7% yield, 80% purity) as a yellow solid. ¹H NMR(400 MHz, DMSO-d₆) δ ppm 9.21 (d, J=1.6 Hz, 1H), 8.77 (d, J=2.0 Hz, 1H),8.62 (s, 1H), 7.99-7.95 (m, 2H), 7.64 (d, J=8.0 Hz, 1H), 7.31 (d, J=8.4Hz, 1H), 7.23 (d, J=2.0 Hz, 1H), 7.09-6.99 (m, 2H), 6.66 (s, 1H), 3.84(q, J=6.0 Hz, 2H), 3.27-3.22 (m, 1H), 3.13-3.10 (m, 2H), 1.35 (d, J=6.8Hz, 6H); ES-LCMS m/z 474.9, 476.9 [M+H]⁺.

Step 5:5-[7-[2-(1H-Indol-3-yl)ethylamino]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile (I-81)

A mixture of5-(5-bromo-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine (70 mg, 117.80 μmol, 1 eq), Pd(PPh₃)₄ (13.61mg, 11.78 μmol, 0.1 eq) and Zn(CN)₂ (27.67 mg, 235.60 μmol, 2 eq) in DMF(5 mL) was degassed and purged with N₂ for 3 times. The mixture wasstirred at 85° C. for 12 h under N₂ atmosphere. The mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Gemini 150*25 5 u; mobile phase: [water(0.05% ammonia hydroxide v/v)-ACN]; B %: 60%-90%, 10 min) to yield5-[7-[2-(1H-indol-3-yl)ethylamino]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-5-yl]pyridine-3-carbonitrile(18.46 mg, 43.72 μmol, 37.1% yield, 99.819% purity) as a yellow solid.¹H NMR (400 MHz, CD₃OD) δ ppm 9.07 (d, J=2.0 Hz, 1H), 8.86 (d, J=1.6 Hz,1H), 7.97 (t, J=2.0 Hz, 1H), 7.90 (s, 1H), 7.74-7.71 (m, 1H), 7.18-7.17(m, 1H), 7.13-7.10 (m, 2H), 6.95 (s, 1H), 5.86 (s, 1H), 3.87 (t, J=6.0Hz, 2H), 3.28-3.19 (m, 1H), 3.18 (t, J=6.0 Hz, 2H), 1.37 (d, J=6.8 Hz,6H); ES-LCMS m/z 422.0 [M+H]⁺.

Example 80

Synthesis of I-82a, I-82b and I-82c

Synthetic Scheme:

Step 1: N-Benzyl-6-methoxy-tetralin-2-amine

To a solution of 6-methoxytetralin-2-one (300 mg, 1.70 mmol, 1 eq) inanhydrous DCM (5 mL) was added phenylmethanamine (182.43 mg, 1.70 mmol,185.58 μL, 1 eq), AcOH (102.23 mg, 1.70 mmol, 97.37 μL, 1 eq) andNaBH(OAc)₃ (541.24 mg, 2.55 mmol, 1.5 eq). The mixture was stirred at25° C. for 12 h. Aq. NaOH (1 M, 20 mL) was added and the mixture wasextracted with DCM (20 mL×3). The combined organic layers were washedwith brine (20 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated to afford the crude product which was purified on silicagel column chromatography (from PE/EtOAc=1/0 to 10/1, TLC: PE/EtOAc=2/1,R_(f)=0.67) to give N-benzyl-6-methoxy-tetralin-2-amine (255 mg, 883.18μmol, 51.88% yield, 92.6% purity) as brown oil. ¹H NMR (400 MHz, CD₃OD)δ ppm 7.38-7.28 (m, 4H), 7.26-7.21 (m, 1H), 6.98 (d, J=8.4 Hz, 1H), 6.67(dd, J=2.5, 8.3 Hz, 1H), 6.61 (d, J=2.4 Hz, 1H), 3.89 (s, 2H), 3.75 (s,3H), 3.04-2.93 (m, 2H), 2.91-2.72 (m, 2H), 2.64-2.50 (m, 1H), 2.12-1.99(m, 1H), 1.62-1.56 (m, 2H); ES-LCMS m/z 268.2 [M+H]⁺.

Step 2: 6-Methoxytetralin-2-amine

To a solution of N-benzyl-6-methoxy-tetralin-2-amine (255 mg, 883.18μmol, 1 eq) in anhydrous EtOH (10 mL) and anhydrous THE (10 mL) wasadded Pd/C (10%, 255 mg) under N₂ atmosphere. The suspension wasdegassed and purged with H₂ for 3 times. The mixture was stirred underH₂ (50 psi) at 50° C. for 8 h. The mixture was filtered and concentratedto afford the crude product of 6-methoxytetralin-2-amine (210 mg, 874.40μmol, 99.0% yield, 73.8% purity) as black oil which was used for thenext step without further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm6.94 (d, J=8.4 Hz, 1H), 6.68-6.64 (m, 1H), 6.62 (d, J=2.4 Hz, 1H),3.74-3.71 (m, 3H), 3.10 (ddt, J=3.3, 5.1, 10.2 Hz, 1H), 2.93 (dd, J=4.1,15.5 Hz, 1H), 2.88-2.81 (m, 2H), 2.49 (dd, J=9.8, 15.5 Hz, 1H),2.06-1.97 (m, 1H), 1.64-1.51 (m, 1H); ES-LCMS m/z 178.3 [M+H]⁺.

Step 3: 2-Aminotetralin-6-ol

A solution of 6-methoxytetralin-2-amine (210 mg, 874.39 μmol, 1 eq) inHBr solution (5 mL) was stirred at 100° C. for 8 h. The mixture wasconcentrated to give the crude product 2-aminotetralin-6-ol (195 mg,350.66 μmol, 40.10% yield, 43.9% purity, HBr) as a brown solid which wasused for the next step without further purification. ¹H NMR (400 MHz,CD₃OD) δ ppm 6.91 (d, J=8.4 Hz, 1H), 6.58 (dd, J=2.4, 8.4 Hz, 1H), 6.53(s, 1H), 3.54-3.44 (m, 1H), 3.06 (dd, J=5.1, 15.4 Hz, 1H), 2.86 (dd,J=5.1, 8.2 Hz, 2H), 2.71 (dd, J=10.1, 15.4 Hz, 1H), 2.21-2.12 (m, 1H),1.85-1.75 (m, 1H); ES-LCMS m/z 164.2 [M+H]⁺.

Step 4:2-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(100 mg, 337.09 μmol, 1 eq) in i-prOH (5 mL) was added DIEA (130.70 mg,1.01 mmol, 176.14 μL, 3 eq) and 2-aminotetralin-6-ol (150.39 mg, 404.51μmol, 1.2 eq). The mixture was stirred at 120° C. for 3 h undermicrowave under N₂ atmosphere. The mixture was concentrated to removethe solvent. H₂O (10 mL) was added, extracted with EtOAc (10 mL×3). Thecombined organic layers were washed with brine (10 mL), dried overanhydrous Na₂SO₄, filtered and concentrated to afford the crude productwhich was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 5/1, TLC: PE/EtOAc=1/1, R_(f)=0.60) to give2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol(78 mg, 184.97 μmol, 54.9% yield, 99.0% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 9.15 (s, 1H), 8.51 (d, J=2.6 Hz, 1H), 8.36(d, J=9.3 Hz, 1H), 7.92 (s, 1H), 6.93 (d, J=7.5 Hz, 1H), 6.74 (s, 1H),6.61-6.53 (m, 2H), 4.58 (s, 2H), 4.35-4.23 (m, 1H), 2.94-2.81 (m, 2H),2.35-2.18 (m, 1H), 2.09-1.86 (m, 2H), 1.40 (d, J=7.1 Hz, 6H); ES-LCMSm/z 418.0 [M+H]⁺.

Step 5:(2R)-2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol(I-82b) &(2S)-2-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol(I-82a)

2-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol(78 mg, 184.97 μmol, 1 eq) was separated by SFC (column: AD (250 mm*30mm, 5 μm); mobile phase: [0.1% NH₃H₂O IPA]; B %: 40%-40%, min). Thesolution after separation were concentrated to afford the crude productswhich were purified by preparative HPLC (MeCN/H₂O as eluents, acidiccondition, Instrument: Phenomenex Synergi C18 150*30 mm*4 um/Mobilephase: water (0.05% HCl)-ACN/Gradient: B from 55% to 85% in 12 min/Flowrate: 25 mL/min), followed by lyophilization to yield an enantiomer(24.75 mg, 50.47 μmol, 27.3% yield, 100% purity, 2HCl, SFC: T_(R)=5.168min, ee=100%, OR: [α]^(26.6) _(D)=167.660) as a yellow solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 8.97 (s, 1H), 8.79 (d, J=2.6 Hz, 1H), 8.34-8.28(m, 1H), 8.25 (s, 1H), 6.98-6.91 (m, 2H), 6.61-6.55 (m, 2H), 4.47 (s,1H), 3.40-3.33 (m, 1H), 3.18 (dd, J=4.6, 15.4 Hz, 1H), 3.09-2.99 (m,2H), 2.96-2.87 (m, 1H), 2.25 (s, 1H), 2.07 (dq, J=5.5, 11.8 Hz, 1H),1.38 (d, J=7.1 Hz, 6H); ES-LCMS m/z 418.0 [M+H]⁺; and the otherenantiomer (26.47 mg, 53.98 μmol, 29.2% yield, 100% purity, 2HCl, SFC:T_(R)=5.742 min, ee=100%, OR: [α]^(26.8) _(D)=−167.394) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.97 (s, 1H), 8.78 (d, J=2.9 Hz,1H), 8.30 (td, J=2.2, 9.0 Hz, 1H), 8.24 (s, 1H), 6.98-6.91 (m, 2H),6.62-6.54 (m, 2H), 4.48 (s, 1H), 3.37-3.31 (m, 1H), 3.22-3.13 (m, 1H),3.09-2.98 (m, 2H), 2.96-2.87 (m, 1H), 2.26 (d, J=10.4 Hz, 1H), 2.07(tdd, J=5.6, 11.6, 17.7 Hz, 1H), 1.38 (d, J=6.8 Hz, 6H); ES-LCMS m/z418.0 [M+H]⁺.

Example 81

Synthesis of I-83

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-83)

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidine(50 mg, 160.79 μmol, 1 eq), 2-(1H-indol-3-yl)ethanamine (25.76 mg,160.79 μmol, 1 eq) and DIEA (62.34 mg, 482.37 μmol, 84.02 μL, 3 eq) ini-PrOH (5 mL) was stirred at 80° C. for 2 h. The mixture wasconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Synergi C18 150*30 mm*4 um;mobile phase: [water(0.05% HCl)-ACN]; B %: 50%-80%, 12 min) to yield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(41.24 mg, 76.67 μmol, 47.7% yield, 100% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.68 (d, J=2.8 Hz, 1H), 8.33 (s,1H), 7.51 (d, J=7.8 Hz, 1H), 7.47-7.45 (m, 1H), 7.15 (d, J=7.6 Hz, 1H),7.02 (s, 1H), 6.97 (t, J=7.6 Hz, 1H), 6.84-6.82 (m, 1H), 5.71 (s, 1H),4.01-3.97 (m, 2H), 3.27 (t, J=7.2 Hz, 1H), 3.23-3.19 (m, 2H), 2.57 (s,3H), 1.36 (d, J=7.2 Hz, 6H); ES-LCMS m/z 429.3 [M+H]⁺.

Example 82

Synthesis of I-84

Synthetic Scheme:

Step 1: 2-Acetyl-3-methyl-butanenitrile

To a mixture of DIPA (1.22 g, 12.03 mmol, 1.70 mL, 1 eq) in THE (10 mL)was added n-BuLi (2.5 M in THF, 5.05 mL, 1.05 eq) dropwise at −78° C.under N₂. The mixture was stirred at −78° C. for 15 min, then warmed upto 0° C. and stirred for 1 h. The mixture was cooled to −78° C.,3-methylbutanenitrile (1 g, 12.03 mmol, 1.27 mL, 1 eq) in THE (10 mL)was added dropwise and stirred at −78° C. for 15 min. A solution ofethyl acetate (1.11 g, 12.63 mmol, 1.24 mL, 1.05 eq) in THF (10 mL) wasadded dropwise. The mixture was stirred at 20° C. for 16 h. TLC(PE/EtOAc=1/1, R_(f)=0.74) indicated one new spot formed. The mixturewas quenched with 2N HCl (10 mL), extracted with EtOAc (50 mL×3), washedwith brine (20 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by flash silicagel chromatography (from PE/EtOAc=10/1 to 1/1, TLC: PE/EtOAc=1/1,R_(f)=0.74) to yield 2-acetyl-3-methyl-butanenitrile (661 mg, 4.22 mmol,35.1% yield, 80.0% purity) as yellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm3.32 (d, J=5.2 Hz, 1H), 2.44-2.38 (m, 1H), 2.37 (s, 3H), 1.11 (d, J=6.8Hz, 3H), 1.09-1.05 (m, 3H); ES-LCMS: No correct mass was found.

Step 2: 4-Isopropyl-3-methyl-1H-pyrazol-5-amine

A mixture of 2-acetyl-3-methyl-butanenitrile (750.00 mg, 4.79 mmol, 1eq), hydrazine hydrate (311.96 mg, 6.23 mmol, 302.87 μL, 1.3 eq) andAcOH (503.76 mg, 8.39 mmol, 479.77 μL, 1.75 eq) in EtOH (20 mL) wasstirred at 90° C. for 16 h under N₂. The mixture was concentrated underreduced pressure to give a residue which was diluted with sat.NaHCO₃ (20mL) and extracted with DCM (20 mL×3). The combined organic layers weredried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to give 4-isopropyl-3-methyl-1H-pyrazol-5-amine (560 mg, 3.62mmol, 75.5% yield, 90.0% purity) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 2.82-2.69 (m, 1H), 2.18 (s, 3H), 1.23 (d, J=6.8 Hz, 6H);ES-LCMS m/z 140.2 [M+H]⁺.

Step 3:5-(5-Fluoro-3-pyridyl)-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidin-7-ol

A mixture of 4-isopropyl-3-methyl-1H-pyrazol-5-amine (502.05 mg, 3.25mmol, 0.8 eq) and methyl 3-(5-fluoro-3-pyridyl)-3-oxo-propanoate (816.33mg, 4.06 mmol, 1 eq) in AcOH (10 mL) was stirred at 120° C. for 3 h. Themixture was concentrated under reduced pressure to give5-(5-fluoro-3-pyridyl)-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidin-7-ol (1.2 g, crude) as a purple solid which was used innext step without further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm8.82 (s, 1H), 8.68-8.66 (m, 1H), 8.14-8.08 (m, 2H), 3.30-3.24 (m, 1H),2.46-2.41 (m, 3H), 1.37 (d, J=7.2 Hz, 6H); ES-LCMS m/z 287.2 [M+H]⁺.

Step 4:7-Chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidine

A solution of5-(5-fluoro-3-pyridyl)-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidin-7-ol(1.2 g, 4.19 mmol, 1 eq) in POCl₃ (16.3 g, 106.31 mmol, 9.88 mL, 25.36eq) was stirred at 110° C. for 2 h. The mixture was concentrated underreduced pressure to give a residue which was purified by flash silicagel chromatography (from PE/EtOAc=10/1 to 1/1, TLC: PE/EtOAc=1/1,R_(f)=0.75) to yield7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidine(381 mg, 1.23 mmol, 29.2% yield, 98.0% purity) as a yellow solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 9.08 (s, 1H), 8.57 (d, J=2.4 Hz, 1H), 8.20-8.15(m, 1H), 7.28 (s, 1H), 3.36-3.24 (m, 1H), 2.57 (s, 3H), 1.48 (d, J=6.8Hz, 6H); ES-LCMS m/z 305.0 [M+H]⁺.

Step 5:N-[2-(5,7-Difluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-84)

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidine(50 mg, 160.79 μmol, 1 eq),2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethanamine (33.80 mg, 160.79μmol, 1 eq) and DIEA (62.34 mg, 482.36 μmol, 84.02 μL, 3 eq) in i-PrOH(10 mL) was stirred at 80° C. for 2 h. The mixture was concentratedunder reduced pressure to give a residue which was purified bypreparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um; mobilephase: [water(0.05% HCl)-ACN]; B %: 58%-78%, 10 min) to yieldN-[2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-2-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(32.01 mg, 54.45 μmol, 33.9% yield, 100% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.75 (d, J=2.8 Hz, 1H), 8.43 (s,1H), 7.66-7.63 (m, 1H), 6.93-6.89 (m, 1H), 6.46-6.39 (m, 1H), 5.73 (s,1H), 3.97-3.94 (m, 2H), 3.31-3.23 (m, 1H), 3.13-3.09 (m, 2H), 2.57 (s,3H), 2.23 (s, 3H), 1.36 (d, J=7.2 Hz, 6H); ES-LCMS m/z 479.3 [M+H]⁺.

Example 83

Synthesis of I-85

Synthetic Scheme:

Step 1:N-(2-(5,7-Difluoro-2-methyl-1H-indol-3-yl)ethyl)-5-(5-fluoropyridin-3-yl)pyrazolo[1,5-a]pyrimidin-7-amine(I-85)

A mixture of 7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(40 mg, 147.68 μmol, 1 eq),2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethanamine (60 mg, 153.73 μmol,1.04 eq, 2 oxalic acid) and DIEA (60.00 mg, 464.24 μmol, 80.86 μL, 3.14eq) in i-PrOH (3 mL) was stirred at 50° C. for 19 h. The mixture wasconcentrated under reduced pressure to dryness to give a residue whichwas purified by preparative HPLC (column: Phenomenex Synergi C18 150×30mm×4 um; mobile phase: [water(0.05% HCl)-ACN]; B %: 30%-52%, 12 min).The desired fraction was lyophilized to giveN-(2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethyl)-5-(5-fluoropyridin-3-yl)pyrazolo[1,5-a]pyrimidin-7-amine(22.94 mg, 43.14 μmol, 29.2% yield, 100.0% purity, 3HCl) as an off-whitesolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.71 (d, J=2.4 Hz, 1H), 8.47 (s,1H), 8.22 (d, J=2.4 Hz, 1H), 7.59 (td, J=2.4, 9.2 Hz, 1H), 7.01 (dd,J=2.4, 9.2 Hz, 1H), 6.51 (d, J=2.4 Hz, 1H), 6.44 (ddd, J=2.4, 9.6, 11.2Hz, 1H), 5.85 (s, 1H), 4.04-3.92 (m, 2H), 3.17-3.08 (m, 2H), 2.13 (s,3H); ES-LCMS m/z 423.2 [M+H]⁺.

Example 84

Synthesis of I-86

Synthetic Scheme:

Step 1: 2-Cyclopropyl-3-oxo-propanenitrile

To a solution of DIPA (3.74 g, 36.98 mmol, 5.23 mL, 1 eq) in THE (15 mL)was added n-BuLi (2.5 M in n-hexane, 15.53 mL, 1.05 eq). The mixture wasstirred at −78° C. for 10 min. The solution of 2-cyclopropylacetonitrile(3 g, 36.98 mmol, 3.42 mL, 1 eq) in THE (15 mL) was added into the abovemixture dropwise. Then the mixture was stirred at −78° C. for 10 minunder N₂ atmosphere. A solution of ethyl formate (2.88 g, 38.88 mmol,3.13 mL, 1.05 eq) in THE (15 mL) was added dropwise and stirred at −78°C. for 40 min. Then the reaction mixture was warmed to 25° C. for 12 h.TLC (PE/EtOAc=1/1, R_(f)=0.50) showed one major new spot was detected.The reaction mixture was quenched by addition 1 N HCl solution (50 mL)at 0° C., extracted with EtOAc (50 mL×3). The combined organic layerswere dried over Na₂SO₄, filtered and concentrated under reduced pressureto give the residue which was purified by flash silica gelchromatography (from PE/EtOAc=100/1 to 1/1, TLC: PE/EtOAc=1/1,R_(f)=0.50) to yield compound 2-cyclopropyl-3-oxo-propanenitrile (3.4 g,crude) as yellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.61 (d, J=1.1 Hz,1H), 4.61-4.55 (m, 1H), 4.01-3.84 (m, 1H), 1.92-1.79 (m, 2H), 1.66-1.52(m, 2H).

Step 2: 4-Cyclopropyl-1H-pyrazol-5-amine

To a solution of 2-cyclopropyl-3-oxo-propanenitrile (3.4 g, 31.16 mmol,1 eq) in EtOH (30 mL) was added AcOH (3.27 g, 54.52 mmol, 3.12 mL, 1.75eq) and hydrazine (1.30 g, 40.50 mmol, 1.46 mL, 1.3 eq). The mixture wasstirred at 90° C. for 12 h. TLC (PE/EtOAc=1/1, R_(f)=0.10) showed onemajor new spot was detected. The reaction mixture was concentrated underreduced pressure to give the residue which was diluted with NaHCO₃solution (200 mL) and extracted with EtOAc (100 mL×3). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated underreduced pressure to give the residue which was purified by flash silicagel chromatography (from DCM/MeOH=100/1 to 10/1, TLC: PE/EtOAc=10/1,R_(f)=0.70) to yield 4-cyclopropyl-1H-pyrazol-5-amine (1.7 g, crude) asa brown solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.04 (d, J=0.7 Hz, 1H),1.49-1.39 (m, 1H), 0.83-0.75 (m, 2H), 0.49-0.41 (m, 2H); ES-LCMS m/z nodesired MS was detected.

Step 3:3-Cyclopropyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol

To a solution of methyl 3-(5-fluoro-3-pyridyl)-3-oxo-propanoate (500 mg,2.49 mmol, 1 eq) in AcOH (6 mL) was added4-cyclopropyl-1H-pyrazol-5-amine (370.54 mg, 3.01 mmol, 1.21 eq). Themixture was stirred at 120° C. for 2 h. The reaction mixture wasconcentrated under reduced pressure to yield3-cyclopropyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol (830mg, crude) as a yellow solid which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.90 (s, 1H), 8.80(d, J=2.6 Hz, 1H), 8.25 (td, J=2.2, 9.7 Hz, 1H), 7.68 (s, 1H), 6.09 (s,1H), 2.05 (m, 2H), 0.94-0.87 (m, 2H), 0.73-0.64 (m, 2H); ES-LCMS m/z271.2 [M+H]⁺.

Step 4:7-Chloro-3-cyclopropyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine

To a solution of3-cyclopropyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol (830mg, 3.07 mmol, 1 eq) in POCl₃ (5 mL) was stirred at 110° C. for 1 h. Thereaction mixture was concentrated under reduced pressure to give theresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=100/1 to 2/1, TLC: PE/EtOAc=3/1, R_(f)=0.40) to yield compound7-chloro-3-cyclopropyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(290 mg, 924.12 μmol, 30.1% yield, 92.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.09 (s, 1H), 8.60 (d, J=2.6 Hz, 1H), 8.20(td, J=2.1, 9.4 Hz, 1H), 8.01 (s, 1H), 7.37 (s, 1H), 2.17 (tt, J=5.2,8.4 Hz, 1H), 1.11-1.03 (m, 2H), 1.02-0.95 (m, 2H); ES-LCMS m/z 289.0,291.0 [M+H]⁺.

Step 5:3-Cyclopropyl-5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-86)

To a solution of7-chloro-3-cyclopropyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(45 mg, 143.40 μmol, 1.0 eq) in i-PrOH (3 mL) was added DIEA (55.60 mg,430.19 μmol, 74.93 μL, 3.0 eq) and 2-(1H-indol-3-yl)ethanamine (29.87mg, 186.42 μmol, 1.3 eq). The mixture was stirred at 50° C. for 3 h. Thereaction mixture was concentrated under reduced pressure to give theresidue which was purified by preparative HPLC (HCl condition, column:Phenomenex Gemini 150×25 mm×10 um; mobile phase: [water(0.05% HCl)-ACN];B %: 43%-73%, 10 min) and the desired fraction was lyophilized to yield3-cyclopropyl-5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(40.50 mg, 77.61 μmol, 54.1% yield, 100% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.73 (d, J=2.5 Hz, 1H), 8.36 (s,1H), 8.00 (s, 1H), 7.54-7.46 (m, 2H), 7.15 (d, J=8.0 Hz, 1H), 7.00 (s,1H), 6.97 (t, J=7.5 Hz, 1H), 6.85-6.80 (m, 1H), 5.78 (s, 1H), 4.07-3.99(m, 2H), 3.26-3.19 (m, 2H), 1.96-1.87 (m, 1H), 1.05-0.97 (m, 2H),0.78-0.70 (m, 2H); ES-LCMS m/z 412.9 [M+H]⁺.

Example 85

Synthesis of I-87

Synthetic Scheme:

Step 1:3-Cyclopropyl-N-(2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethyl)-5-(5-fluoropyridin-3-yl)pyrazolo[1,5-a]pyrimidin-7-amine(I-87)

A mixture of7-chloro-3-cyclopropyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(40 mg, 127.46 μmol, 1 eq),2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethanamine (54.72 mg, 140.21μmol, 1.1 eq, 2 oxalic acid) and DIEA (0.055 g, 425.56 μmol, 74.12 μL,3.34 eq) in i-PrOH (3 mL) was stirred at 50° C. for 19 h. The mixturewas concentrated under reduced pressure to dryness to give a residuewhich was purified by preparative HPLC (column: Phenomenex Synergi C18150×30 mm×4 um; mobile phase: [water(0.05% HCl)-ACN]; B %: 45%-75%, 12min). The desired fraction was lyophilized to give3-cyclopropyl-N-(2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethyl)-5-(5-fluoropyridin-3-yl)pyrazolo[1,5-a]pyrimidin-7-amine(22.33 mg, 37.39 μmol, 29.3% yield, 95.7% purity, 3HCl) as an off-whitesolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.73 (d, J=1.6 Hz, 1H), 8.43 (s,1H), 7.97 (s, 1H), 7.62 (d, J=8.8 Hz, 1H), 6.93 (dd, J=2.0, 9.2 Hz, 1H),6.49-6.35 (m, 1H), 5.73 (s, 1H), 3.96 (t, J=5.2 Hz, 2H), 3.10 (t, J=5.6Hz, 2H), 2.14 (s, 3H), 1.98-1.86 (m, 1H), 1.07-0.94 (m, 2H), 0.79-0.67(m, 2H); ES-LCMS m/z 463.2 [M+H]⁺.

Example 86

Synthesis of I-88

Synthetic Scheme:

Step 1:N-[2-(5,7-Difluoro-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidin-4-amine(I-88)

4-Chloro-2-(5-fluoro-3-pyridyl)-6-isopropooxy-pyrimidine (55 mg, 172.59μmol, 1 eq), 2-(5,7-difluoro-1H-indol-3-yl)ethanamine (50.31 mg, 225.64μmol, 1.31 eq) and DIEA (66.92 mg, 517.77 μmol, 90.19 μL, 3.0 eq) ini-PrOH (3 mL) were taken up into a microwave tube. The sealed tube washeated at 125° C. for 6 h under microwave. The reaction mixture wasconcentrated under reduced pressure to give the residue which waspurified by preparative HPLC (HCl condition; column: Phenomenex SynergiC18 150×30 mm×4 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 65%-95%,12 min). The desired fraction was lyophilized to yieldN-[2-(5,7-difluoro-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidin-4-amine(15.59 mg, 28.70 μmol, 16.6% yield, 98.81% purity, 3HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.12 (s, 1H), 8.76 (d, J=2.5Hz, 1H), 8.36 (d, J=8.8 Hz, 1H), 7.21 (s, 1H), 7.11 (dd, J=1.6, 9.2 Hz,1H), 6.66 (t, J=9.9 Hz, 1H), 5.80 (s, 1H), 5.07 (s, 1H), 3.81 (s, 2H),3.06 (t, J=6.7 Hz, 2H), 1.39 (d, J=6.0 Hz, 6H); ES-LCMS m/z 428.2[M+H]⁺.

Example 87

Synthesis of I-89

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[2-(2-methyl-1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amin(I-89)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(40 mg, 132.08 μmol, 1 eq) and 2-(2-methyl-1H-indol-3-yl)ethanamine(34.52 mg, 198.13 μmol, 1.5 eq) in i-PrOH (10 mL) was added DIEA (51.21mg, 396.25 μmol, 69.02 μL, 3.0 eq). The mixture was stirred at 70′C for16 h. The reaction mixture concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: PhenomenexSynergi C18 150*30 mm*4 um; mobile phase: [water(0.05% HCl)-ACN]; B %:47%-72%, 12 min) followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-3-isopropyl-N-[2-(2-methyl-1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(47.07 mg, 86.87 μmol, 65.8% yield, 99.3% purity, 3HCl) as a red solid.¹H NMR (400 MHz, CD₃OD) δ ppm 8.68 (d, J=2.0 Hz, 1H), 8.20 (s, 1H), 8.07(s, 1H), 7.37 (d, J=8.6 Hz, 1H), 7.32-7.27 (m, 1H), 6.86-6.82 (m, 1H),6.69 (dquin, J=1.3, 7.0 Hz, 2H), 5.43 (s, 1H), 3.89-3.83 (m, 2H),3.15-3.07 (m, 1H), 3.03-2.98 (m, 2H), 1.95 (s, 3H), 1.19 (d, J=6.8 Hz,6H); ES-LCMS m/z 429.3 [M+H]⁺.

Example 88

Synthesis of I-90

Synthetic Scheme:

Step 1:3-tert-Butyl-5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-90)

To a solution of3-tert-butyl-7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(40 mg, 128.63 μmol, 1 eq) and 2-(1H-indol-3-yl)ethanamine (30.91 mg,192.95 μmol, 1.5 eq) in i-PrOH (10 mL) was added DIEA (49.87 mg, 385.89μmol, 67.21 μL, 3.0 eq). The mixture was stirred at 70° C. for 16 h. Thereaction mixture was concentrated under reduced pressure to removesolvent to yield a residue which was purified by preparative HPLC(column: Phenomenex Synergi C18 150*30 mm*4 um; mobile phase:[water(0.05% HCl)-ACN]; B %: 70%-90%, 12 min), followed bylyophilization to yield3-tert-butyl-5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(42.66 mg, 78.86 μmol, 61.3% yield, 99.4% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.82 (br s, 1H), 9.13 (s, 1H),8.67 (d, J=2.2 Hz, 1H), 8.22 (d, J=8.6 Hz, 1H), 8.04 (br s, 1H), 7.94(s, 1H), 7.64 (d, J=7.9 Hz, 1H), 7.31 (d, J=7.9 Hz, 1H), 7.23 (d, J=2.0Hz, 1H), 7.10-6.95 (m, 2H), 6.64 (s, 1H), 3.83 (q, J=6.9 Hz, 2H), 3.11(t, J=7.1 Hz, 2H), 1.46 (s, 9H); ES-LCMS m/z 429.3 [M+H]⁺.

Example 89

Synthesis of I-91

Step 1: 2-Formyl-3,3-dimethyl-butanenitrile

To a mixture of DIPA (833.20 mg, 8.23 mmol, 1.16 mL, 1 eq) in TH (20 mL)was added n-BuLi (2.5 M, 3.46 mL, 1.05 eq) dropwise at −78° C. under N₂.The mixture was stirred at −78° C. for 10 min, then heated to 0° C. andstirred for 1 h. The mixture was cooled to −78° C.,3,3-dimethylbutanenitrile (800 mg, 8.23 mmol, 12.66 mL, 1 eq) dissolvedin THE (15 mL, anhydrous) was added dropwise and stirred at −78° C. for10 min. A solution of ethyl formate (640.46 mg, 8.65 mmol, 695.40 μL,1.05 eq) in THF (15 mL, anhydrous) was added dropwise and stirred at−78° C. for 40 min, then the mixture was warmed to 5-14° C. for 16 h.TLC (PE/EtOAc=3/1, R_(f)=0.34) indicated one major new spot wasdetected. The reaction mixture was quenched by addition 1N HCl solution(50) mL at −78° C., extracted with DCM (50 mL×3). The combined organiclayers were dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified on silica gelcolumn chromatography (from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1,R_(f)=0.34) to give the product 2-formyl-3,3-dimethyl-butanenitrile (750mg, 5.39 mmol, 65.5% yield, 90.0% purity) as a yellow oil. ¹H NMR (400MHz, CDCl₃) δ ppm 9.66 (d, J=2.2 Hz, 1H), 3.27 (d, J=2.4 Hz, 1H), 1.23(s, 9H).

Step 2: 4-tert-Butyl-1H-pyrazol-5-amine

A mixture of 2-formyl-3,3-dimethyl-butanenitrile (750 mg, 5.39 mmol, 1eq), hydrazine (224.65 mg, 7.01 mmol, 253.56 μL, 1.3 eq) and AcOH(566.71 mg, 9.44 mmol, 539.73 μL, 1.75 eq) in EtOH (20 mL) was degassedand purged with N₂ for 3 times. Then the mixture was stirred at 80-90°C. (reflux) for 16 h under N₂ atmosphere. TLC (PE/EtOAc=1/1, R_(f)=0.55)indicated most of starting material was consumed and one major new spotwith larger polarity was detected. The reaction mixture concentratedunder reduced pressure. The residue was diluted with H₂O (20 mL),adjusted to pH to 9-10 with NaHCO₃ solid and extracted with DCM (20mL×3). The combined organic layers were dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give the crudeproduct 4-tert-butyl-1H-pyrazol-5-amine (750 mg, 5.39 mmol, 99.9% yield,crude purity) as a yellow solid which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.12 (s, 1H), 1.30(s, 9H).

Step 3: 5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-ol

To a solution of methyl 3-(5-fluoro-3-pyridyl)-3-oxo-propanoate (1.08 g,5.37 mmol, 1 eq) in AcOH (20 mL) was added4-tert-butyl-1H-pyrazol-5-amine (747.23 mg, 5.37 mmol, 1 eq). Themixture was stirred at 120° C. for 0.5 h. The reaction mixture wasconcentrated under reduced pressure to remove solvent to give crudeproduct 5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-ol(1.46 g, 5.36 mmol, 100.0% yield, crude) as a yellow oil which was usedin the next step without further purification.

Step 4:3-tert-Butyl-7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine

A mixture of3-tert-butyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol (1.67g, 5.83 mmol, 1 eq) in POCl₃ (37.81 g, 246.59 mmol, 22.92 mL, 42.28 eq)was degassed and purged with N₂ for 3 times. Then the mixture wasstirred at 110° C. for 3 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to remove solvent. The residue wasdiluted with DCM (50 mL×2) and concentrated under reduced pressure togive a residue which was purified on silica gel column chromatography(from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1, R_(f)=0.48) to give theproduct3-tert-butyl-7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(950 mg, 3.05 mmol, 52.4% yield, 98.0% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.13 (t, J=1.5 Hz, 1H), 8.61 (d, J=2.8 Hz, 1H),8.22-8.16 (m, 1H), 8.12 (s, 1H), 7.41 (s, 1H), 1.56 (s, 9H); ES-LCMS m/z305.1, 307.1 [M+H]⁺.

Step 5:3-tert-Butyl-N-[2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-amine(I-91)

To a solution of3-tert-butyl-7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(40 mg, 128.63 μmol, 1 eq) and2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethanamine (57.93 mg, 192.95μmol, 1.5 eq, oxalic acid) in i-PrOH (10 mL) was added DIEA (49.87 mg,385.89 μmol, 67.21 μL, 3.0 eq). The mixture was stirred at 70° C. for 16h. The reaction mixture concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: PhenomenexSynergi C18 150*30 mm*4 um; mobile phase: [water(0.05% HCl)-ACN]; B %:70%-90%, 12 min)3-tert-butyl-N-[2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-amine(34.14 mg, 57.39 μmol, 44.6% yield, 98.8% purity, 3HCl) as a orangesolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.21 (s, 1H), 9.06-8.98 (m, 1H),8.66 (d, J=2.6 Hz, 1H), 8.13-8.01 (m, 2H), 7.95 (s, 1H), 7.22 (dd,J=2.1, 9.6 Hz, 1H), 6.76 (ddd, J=2.2, 9.6, 11.4 Hz, 1H), 6.42 (s, 1H),3.71 (q, J=6.8 Hz, 2H), 3.01 (t, J=6.6 Hz, 2H), 2.20 (s, 3H), 1.47 (s,9H); ES-LCMS m/z 479.3 [M+H]⁺.

Example 90

Synthesis of I-92

Synthetic Scheme:

Step 1: Methyl 5-amino-1H-pyrazole-4-carboxylate

A solution of 5-amino-1H-pyrazole-4-carboxylic acid (1 g, 7.87 mmol, 1eq) in HCl/MeOH (4 M, 40 mL, 20.34 eq) was stirred at 60° C. for 12 hr.The reaction mixture was concentrated under reduced pressure to givemethyl 5-amino-1H-pyrazole-4-carboxylate (1.4 g, 7.49 mmol, 95.2% yield,95.0% purity, HCl) as a white solid which was used in the next stepwithout further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.31 (s,1H), 3.84 (s, 3H); ES-LCMS m/z 142.2 [M+H]⁺.

Step 2: Methyl5-(5-fluoro-3-pyridyl)-7-hydroxy-pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of methyl 3-(5-fluoro-3-pyridyl)-3-oxo-propanoate (160 mg,795.28 μmol, 1 eq) in AcOH (5 mL) was added methyl5-amino-1H-pyrazole-4-carboxylate (148.67 mg, 795.28 μmol, 1 eq, HCl).The mixture was stirred at 120° C. for 4 h. The reaction mixture wasconcentrated to yield methyl5-(5-fluoro-3-pyridyl)-7-hydroxy-pyrazolo[1,5-a]pyrimidine-3-carboxylate(230 mg, crude) as a yellow solid which was used in the next stepwithout further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.83 (s,1H), 8.72 (d, J=2.6 Hz, 1H), 8.29 (s, 1H), 8.22 (s, 1H), 6.34 (s, 1H),3.92 (s, 3H); ES-LCMS m/z 289.2 [M+H]⁺.

Step 3: Methyl7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

A solution of methyl5-(5-fluoro-3-pyridyl)-7-hydroxy-pyrazolo[1,5-a]pyrimidine-3-carboxylate(230 mg, 797.96 μmol, 1 eq) in POCl₃ (5 mL) was stirred at 110° C. for 1h. The reaction mixture was concentrated under reduced pressure to givethe residue which was purified by flash silica gel chromatography(ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0˜50% Ethylacetate/Petroleum ethergradient @ 30 mL/min) to yield methyl7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(180 mg, 516.50 μmol, 64.7% yield, 88.0% purity) as a white solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.08 (s, 1H), 8.65 (s, 1H), 8.59 (d, J=2.6Hz, 1H), 7.69 (s, 1H), 7.56 (s, 1H), 3.93 (s, 3H); ES-LCMS m/z 307.1,309.1 [M+H]⁺.

Step 4: Methyl5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of methyl7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(180 mg, 516.50 μmol, 1 eq), 2-(1H-indol-3-yl)ethanamine (82.75 mg,516.50 μmol, 1 eq) in i-PrOH (10 mL) was added DIEA (200.26 mg, 1.55mmol, 269.90 μL, 3 eq). Then the mixture was stirred at 60° C. for 3 h.The reaction mixture was concentrated under reduced pressure to givemethyl5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylate(120 mg, 242.55 μmol, 47.0% yield, 87.0% purity) as an off white solidwhich was used in the next step without further purification. ¹H NMR(400 MHz, CD₃OD) δ ppm 8.59 (s, 1H), 8.46 (d, J=2.9 Hz, 1H), 8.41 (s,1H), 7.84 (br s, 1H), 7.15-7.15 (m, 1H), 7.13-7.08 (m, 3H), 6.94 (s,1H), 6.07 (s, 1H), 3.86 (s, 3H), 3.11-3.05 (m, 4H); ES-LCMS m/z 431.1[M+H]⁺.

Step 5:5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To a solution of methyl5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylate(60 mg, 121.27 μmol, 1 eq) in MeOH (2 mL) THF (2 mL) and Water (4 mL)was added LiOH H₂O (72.00 mg, 1.72 mmol, 14.15 eq) and the mixture wasstirred at 50° C. for 1 h. The reaction mixture was concentrated underreduced pressure to give a residue which diluted with EtOAc (20 mL) andextracted with EtOAc (20 mL×2). The combined organic layers were washedwith brine (20 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (50 mg, 84.05 μmol, 69.3% yield, 70.0% purity) as a brown solidwhich was used in the next step without further purification. ¹H NMR(400 MHz, CD₃OD) ppm 8.58 (s, 1H), 8.46 (d, J=2.4 Hz, 1H), 8.40 (s, 1H),7.62 (s, 1H), 7.54 (d, J=7.7 Hz, 1H), 7.35 (d, J=8.2 Hz, 1H), 7.10 (d,J=7.9 Hz, 1H), 6.95 (m, 2H), 6.07 (s, 1H), 3.90 (t, J=6.2 Hz, 2H),3.13-3.08 (m, 2H); ES-LCMS m/z 417.2 [M+H]⁺.

Step 6:5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]-N-methoxy-N-methyl-pyrazolo[1,5-a]pyrimidine-3-carboxamide(I-92)

To a solution of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (50 mg, 84.05 μmol, 1 eq) and N-methoxymethanamine (12.30 mg,126.08 μmol, 1.5 eq, HCl) in DCM (5 mL) was added HATU (47.94 mg, 126.08μmol, 1.5 eq) and TEA (17.01 mg, 168.10 μmol, 23.40 μL, 2 eq). Then themixture was stirred at 20° C. for 1 h. The reaction mixture wasconcentrated under reduced pressure to give the residue which wasdiluted with EtOAc (20 mL) and extracted with EtOAc (20 mL×2). Thecombined organic layers were washed with brine (20 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: PhenomenexGemini 150*25 mm*10 um; mobile phase: [water(0.05% HCl)-ACN]; B %:30%-60%, 10 min). The desired fraction was evaporated to afford5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]-N-methoxy-N-methyl-pyrazolo[1,5-a]pyrimidine-3-carboxamide(20.19 mg, 35.14 μmol, 41.8% yield, 99.0% purity, 3HCl) as a lightyellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.74 (d, J=2.6 Hz, 1H), 8.61(s, 1H), 8.40 (s, 1H), 7.57-7.51 (m, 2H), 7.10 (d, J=8.4 Hz, 1H), 7.01(s, 1H), 6.94 (t, J=7.3 Hz, 1H), 6.87-6.81 (m, 1H), 6.04 (s, 1H), 4.07(t, J=5.8 Hz, 2H), 3.85 (s, 3H), 3.37 (s, 3H), 3.24 (t, J=5.7 Hz, 2H);ES-LCMS m/z 460.4 [M+H]⁺.

Example 91

Synthesis of I-93

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-N6-[2-(1H-indol-3-yl)ethyl]-N4-isopropyl-N4-methyl-pyrimidine-4,6-diamine(I-93)

6-Chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine(60 mg, 163.13 μmol, 1.0 eq), N-methylpropan-2-amine (702.00 mg, 9.60mmol, 1 mL, 58.84 eq) and DIEA (63.25 mg, 489.39 μmol, 85.24 μL, 3.0 eq)in i-PrOH (2 mL) were taken up into a microwave tube. The sealed tubewas heated at 125° C. for 10 h under microwave. The reaction mixture wasconcentrated under reduced pressure to give the residue which waspurified by preparative HPLC (HCl condition; column: column: PhenomenexSynergi C18 150×30 mm×4 um; mobile phase: [water(0.05% HCl)-ACN]; B %:22%-52%, 12 min) and the desired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N6-[2-(1H-indol-3-yl)ethyl]-N4-isopropyl-N4-methyl-pyrimidine-4,6-diamine(19.96 mg, 49.25 μmol, 30.2% yield, 99.80% purity, 3 HCl salt) as ayellow solid. ¹H NMR (400 MHz, CD₃OD+Na₂CO₃) δ ppm 9.26 (s, 1H), 8.47(d, J=2.6 Hz, 1H), 8.39-8.33 (m, 1H), 7.65-7.59 (m, 1H), 7.35-7.31 (m,1H), 7.12-7.06 (m, 2H), 7.04-6.98 (m, 1H), 5.34 (s, 1H), 4.95 (s, 1H),3.66 (t, J=6.9 Hz, 2H), 3.07 (t, J=6.9 Hz, 2H), 2.79 (s, 3H), 1.18 (d,J=6.8 Hz, 6H); ES-LCMS m/z 405.3 [M+H]⁺.

Example 92

Synthesis of I-94a

Synthesis Scheme:

Step 1: 3-Aminotetralin-6-ol

To a solution of 7-hydroxytetralin-2-one (500 mg, 3.08 mmol, 1 eq) inanhydrous MeOH (5 mL) was added NH₄OAc (7.13 g, 92.49 mmol, 30 eq).After stirring for 5 h, NaBH₄ (349.88 mg, 9.25 mmol, 3 eq) was added.The mixture was stirred at 25° C. for 3 h. The mixture was concentratedto remove the solvent. H₂O (10 mL) was added, the mixture was extractedwith EtOAc (20 mL×2). The aqueous was adjusted by 3 N HCl to pH=6-7,extracted with EtOAc (10 mL×2). The aqueous was adjusted by 1 N NaOH topH=6-7. Then the mixture was lyophilized to give the solid. The solidwas dissolved in i-PrOH (10 mL). The resulting mixture was filtered andconcentrated to give 3-aminotetralin-6-ol (550 mg, 2.66 mmol, 86.4%yield, 79.0% purity) as a yellow oil which was used in the next stepwithout further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm 6.91 (d,J=8.2 Hz, 1H), 6.58 (dd, J=2.4, 8.4 Hz, 1H), 6.51 (s, 1H), 3.53-3.42 (m,1H), 3.06 (dd, J=5.4, 15.5 Hz, 1H), 2.89-2.73 (m, 3H), 2.17 (d, J=10.1Hz, 1H), 1.84-1.74 (m, 1H); ES-LCMS m/z 164.3 [M+H]⁺.

Step 2:3-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(100 mg, 330.21 μmol, 1 eq) in i-PrOH (10 mL) was added DIEA (128.03 mg,990.63 μmol, 172.55 μL, 3 eq) and 3-aminotetralin-6-ol (204.67 mg,990.63 μmol, 3 eq). The mixture was stirred at 120° C. for 3 h undermicrowave under N₂ atmosphere. The mixture was concentrated to removethe solvent. H₂O (10 mL) was added, the mixture was extracted with EtOAc(10 mL×3). The combined organic layers were washed with brine (10 mL),dried over anhydrous Na₂SO₄, filtered and concentrated to afford thecrude product which was purified on silica gel column chromatography(from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=2/1, R_(f)=0.50) to give3-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol(63 mg, 138.83 μmol, 42.0% yield, 92.0% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 9.19 (s, 1H), 8.56 (d, J=2.5 Hz, 1H),8.45-8.37 (m, 1H), 7.96 (s, 1H), 6.98 (d, J=8.3 Hz, 1H), 6.78 (s, 1H),6.66-6.56 (m, 2H), 4.62 (s, 2H), 4.32 (s, 1H), 3.01-2.89 (m, 3H), 2.28(s, 1H), 2.03 (d, J=5.5 Hz, 1H), 1.44 (d, J=7.0 Hz, 6H); ES-LCMS m/z418.2 [M+H]⁺.

Step 3:(3R)-3-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol I-94)

3-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol(100 mg, 220.37 μmol, 1 eq) was separation by SFC (column: Chiralcel OD250*30 5 u; mobile phase: [0.1% NH₃.H₂O EtOH]; B %: 45%-45%, min) toyield the product (Rt=3.922 min) which was purified by preparative HPLC(MeCN/H₂O as eluents, acidic condition, Instrument: Phenomenex Gemini150*25 mm*10 um/Mobile phase: water (0.05% HCl)-ACN/Gradient: B from 50%to 80% in 10 min/Flow rate: 25 mL/min) followed by lyophilization toyield(3R)-3-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]tetralin-6-ol(19.02 mg, 45.10 μmol, 20.47% yield, 99.0% purity) ([α]²⁵ _(D)=−23.996)as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.99 (s, 1H), 8.81 (d,J=2.4 Hz, 1H), 8.38-8.31 (m, 1H), 8.25 (s, 1H), 7.00-6.91 (m, 2H), 6.60(dd, J=2.5, 8.3 Hz, 1H), 6.55 (d, J=2.4 Hz, 1H), 4.50 (s, 1H), 3.41-3.32(m, 1H), 3.23-3.15 (m, 1H), 3.11-3.02 (m, 1H), 3.01-2.94 (m, 1H),2.94-2.86 (m, 1H), 2.32-2.21 (m, 1H), 2.14-2.02 (m, 1H), 1.38 (d, J=6.8Hz, 6H); ES-LCMS m/z 418.2 [M+H]⁺.

Example 93

Synthesis of I-95

Synthetic Scheme:

Step 1: Methyl5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxylate

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(270 mg, 659.39 μmol, 1 eq, 71.0%) in DMF (10 mL) and MeOH (8 mL) wasadded Et₃N (333.62 mg, 3.30 mmol, 458.90 μL, 5.0 eq), Pd(OAc)₂ (22.21mg, 98.91 μmol, 0.15 eq) and DPPF (54.83 mg, 98.91 μmol, 0.15 eq). Themixture was purged with CO (50 psi) three times and stirred at 70° C.for 24 hr under CO (50 psi). The reaction mixture concentrated underhigh vacuum to give a residue which was purified on silica gel columnchromatography (from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1,R_(f)=0.47) to give the product methyl5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxylate(180 mg, 538.32 μmol, 81.6% yield, 94.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.17 (d, J=6.8 Hz, 1H), 8.62 (d, J=5.3 Hz,1H), 8.22 (dd, J=7.9, 17.9 Hz, 2H), 7.81 (d, J=8.2 Hz, 1H), 4.22-4.00(m, 3H), 3.47-3.45 (m, 1H), 1.47 (t, J=7.4 Hz, 6H); ES-LCMS m/z 315.3[M+H]⁺.

Step 2:5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxylicacid

To a solution of methyl5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxylate(180 mg, 538.32 μmol, 1 eq) in MeOH (10 mL) and THE (10 mL), H₂O (5 mL)was added LiOH (128.92 mg, 5.38 mmol, 10 eq). The mixture was stirred at50° C. for 2 h. The reaction mixture was concentrated under reducedpressure. The residue was diluted with H₂O (20 mL), adjusted to pH=3-4with 1N HCl solution, extracted with DCM (20 mL×3). The combined organiclayers were dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give the crude product5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxylicacid (100 mg, 293.05 μmol, 54.4% yield, 88.0% purity) as a yellow solidwhich was used in the next step without further purification. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.23 (s, 1H), 8.65 (d, J=2.6 Hz, 1H), 8.26 (td,J=2.3, 9.3 Hz, 1H), 8.22 (s, 1H), 8.14 (s, 1H), 3.59-3.35 (m, 1H), 1.49(d, J=6.8 Hz, 6H); ES-LCMS m/z 301.1 [M+H]⁺.

Step 3:5-(5-Fluoro-3-pyridyl)-N-[(3-hydroxyphenyl)methyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxamide(I-95)

To a solution of5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxylicacid (70 mg, 205.14 μmol, 1 eq) and 3-(aminomethyl)phenol (50.53 mg,410.27 μmol, 2.0 eq) in DCM (20 mL) was added HATU (117.00 mg, 307.71μmol, 1.5 eq) and DIEA (79.54 mg, 615.41 μmol, 107.19 μL, 3.0 eq). Themixture was stirred at 25° C. for 16 h. The combined reaction mixturewas diluted with H₂O (20 mL), extracted with EtOAc (20 mL×3). Thecombined organic layers were dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Synergi C18 150*30 mm*4 um;mobile phase: [water(0.05% HCl)-ACN]; B %: 47%-77%, 12 min), followed bylyophilization to yield5-(5-fluoro-3-pyridyl)-N-[(3-hydroxyphenyl)methyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxamide(16.16 mg, 33.78 μmol, 16.5% yield, 100.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.41 (t, J=5.8 Hz, 1H),9.34-9.27 (m, 1H), 8.76 (d, J=2.9 Hz, 1H), 8.62-8.49 (m, 1H), 8.35 (s,1H), 8.20 (s, 1H), 7.15 (t, J=7.7 Hz, 1H), 6.88-6.80 (m, 2H), 6.72-6.61(m, 1H), 4.62 (d, J=6.0 Hz, 2H), 3.42-3.35 (m, 1H), 1.41 (d, J=7.1 Hz,6H); ES-LCMS m/z 406.2 [M+H]⁺.

Example 94

Synthesis of I-96

Synthetic Scheme:

Step 1:N-[2-(5,7-Difluoro-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-96)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(40 mg, 137.12 μmol, 1 eq), 2-(5,7-difluoro-1H-indol-3-yl)ethanamine(45.86 mg, 205.68 μmol, 1.5 eq) and DIEA (17.72 mg, 137.12 μmol, 23.88μL, 1 eq) in i-PrOH (10 mL) was stirred at 80° C. for 2 h. The mixturewas concentrated under reduced pressure to give a residue which waswashed with MeOH (10 mL), filtered and dried under reduced pressure toyieldN-[2-(5,7-difluoro-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-amine(18.95 mg, 41.98 μmol, 30.6% yield, 100% purity) as a white solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 11.37 (s, 1H), 9.29 (t, J=1.6 Hz, 1H),9.03-9.01 (m, 1H), 8.70 (d, J=2.8 Hz, 1H), 8.26-8.22 (m, 1H), 8.10 (s,1H), 7.34 (d, J=2.0 Hz, 1H), 7.27 (dd, J=2.0, 7.2 Hz, 1H), 6.91-6.87 (m,1H), 3.92 (q, J=6.8 Hz, 2H), 3.22-3.15 (m, 1H), 3.09 (t, J=7.2 Hz, 2H),1.35 (d, J=6.8 Hz, 6H); ES-LCMS m/z 452.2 [M+H]⁺.

Example 95

Synthesis of I-97a

Synthetic Scheme:

Step 1: 1-Methoxy-4-[(E)-2-nitroprop-1-enyl]benzene

A mixture of 4-methoxybenzaldehyde (5 g, 36.72 mmol, 4.46 mL, 1 eq),NH₄OAc (566.17 mg, 7.34 mmol, 0.2 eq) in EtNO₂ (50 mL) was stirred at110° C. for 5 h under N₂ atmosphere. Then the mixture was stirred at110° C. for another 40 h. TLC (PE/EtOAc=5/1, R_(f)=0.73) indicatedstarting material was consumed completely and one new spot formed. Themixture was concentrated under reduced pressure to give a residue whichwas purified by flash silica gel chromatography (from PE/EtOAc=10/1 to5/1, TLC: PE/EtOAc=5/1, R_(f)=0.73) to yield1-methoxy-4-[(E)-2-nitroprop-1-enyl]benzene (3.7 g, 18.19 mmol, 49.5%yield, 95% purity) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.06(s, 1H), 7.52-7.49 (m, 2H), 7.05-7.02 (m, 2H), 3.85 (s, 3H), 2.45 (s,3H); ES-LCMS m/z 194.0 [M+H]⁺.

Step 2: 1-(4-Methoxyphenyl)propan-2-amine

A mixture of 1-methoxy-4-[(E)-2-nitroprop-1-enyl]benzene (1 g, 4.92mmol, 1 eq) and LAH (1 M in THF, 14.75 mL, 3 eq) in THF (10 mL) wasstirred at 60° C. for 14 h. The mixture was quenched with water (0.5mL), 15% NaOH (0.5 mL) and water (1.5 mL), the mixture was stirred for 3h at 20° C. The mixture was filtered and concentrated under reducedpressure to yield 1-(4-methoxyphenyl)propan-2-amine (800 mg, crude) asyellow oil. ¹H NMR (400 MHz, CD₃OD) δ ppm 7.08 (d, J=8.4 Hz, 2H),6.86-6.81 (m, 2H), 3.72 (s, 3H), 2.96-2.91 (m, 1H), 2.51-2.42 (m, 2H),0.92 (d, J=6.4 Hz, 3H); ES-LCMS m/z 166.2 [M+H]⁺.

Step 3: 4-(2-Aminopropyl)phenol

A mixture of 1-(4-methoxyphenyl) propan-2-amine (1.2 g, 7.26 mmol, 1 eq)in a solution of HBr (8 mL) solution in H₂O (3 mL) was stirred at 100°C. for 3 h. The mixture was extracted with EtOAc (20 mL), pH wasadjusted to 9 with 15% NaOH, the aqueous solution was extracted withEtOAc (20 mL×3), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give 4-(2-aminopropyl) phenol (445 mg, crude) as abrown solid which was used in the next step without furtherpurification. ¹H NMR (400 MHz, CD₃OD) δ ppm 7.00 (d, J=8.4 Hz, 2H),6.73-6.70 (m, 2H), 3.07-3.01 (m, 1H), 2.56-2.51 (m, 2H), 1.07 (d, J=6.4Hz, 3H); ES-LCMS m/z 152.2 [M+H]⁺.

Step 4:4-[(2R)-2-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]propyl]phenol(I-97)

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(100 mg, 330.21 μmol, 1 eq), 4-(2-aminopropyl)phenol (99.86 mg, 660.42μmol, 2 eq) and DIEA (128.03 mg, 990.63 μmol, 172.55 μL, 3 eq) in i-PrOH(10 mL) was degassed and purged with N₂ for 3 times, the mixture wasstirred at 80° C. for 12 h under N₂ atmosphere. The mixture wasconcentrated under reduced pressure to give a residue which was purifiedby SFC (column: Chiralpak AS-H 250*30 5 u; mobile phase: [0.1% NH₃H₂OEtOH]; B %: 20%-20%, min) to give4-[(2R)-2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]propyl]phenol(Rt=4.781 min, 50 mg) and preparative HPLC (column: Phenomenex SynergiC18 150*30 mm*4 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 52%-62%,12 min) to yield4-[(2R)-2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]propyl]phenol(27.45 mg, 57.15 umol, 17.3% yield, 99.6% purity, 2HCl) ([α]²⁵_(D)=−90.432) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.81-8.76(m, 2H), 8.25 (s, 1H), 8.12-8.07 (m, 1H), 7.04 (d, J=8.4 Hz, 2H), 6.54(d, J=8.4 Hz, 2H), 6.34 (s, 1H), 4.42 (br s, 1H), 3.31-3.29 (m, 1H),3.04 (dd, J=4.4, 13.6 Hz, 1H), 2.87-2.83 (m, 1H), 1.54 (d, J=6.4 Hz,3H), 1.36 (t, J=6.8 Hz, 6H); ES-LCMS m/z 406.2 [M+H]⁺.

Example 96

Synthesis of I-98a

Synthetic Scheme:

Step 1: 3-[(E)-2-Nitroprop-1-enyl]-1H-indole

A mixture of 1H-indole-3-carbaldehyde (5 g, 34.45 mmol, 1 eq), NH₄OAc(531.02 mg, 6.89 mmol, 0.2 eq) in nitroethane (50 mL) was stirred at110° C. for 24 h. TLC (DCM/MeOH=10/1, R_(f)=0.8) indicated startingmaterial was consumed completely and one new spot formed. The mixturewas cooled to 0° C. The mixture was filtered and the filtered cake wasdried under reduced pressure to give3-[(E)-2-nitroprop-1-enyl]-1H-indole (5.1 g, 25.22 mmol, 73.2% yield,100% purity) as a brown solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.53 (s,1H), 7.79-7.75 (m, 2H), 7.46 (d, J=8.0 Hz, 1H), 7.26-7.20 (m, 2H), 2.52(s, 3H); ES-LCMS m/z 203.0 [M+H]⁺.

Step 2: 1-(1H-Indol-3-yl)propan-2-amine

To a solution of 3-[(E)-2-nitroprop-1-enyl]-1H-indole (1 g, 4.95 mmol, 1eq) in THE (10 mL) was added LAH (1 M in THF, 14.84 mL, 3 eq) at 0° C.Then the mixture was stirred at 60° C. for 2 h. The mixture was dilutedwith EtOAc (200 mL), quenched with water (0.5 mL), 15% NaOH (0.5 mL),water (1.5 mL), stirred for 3 h, filtered and concentrated under reducedpressure to give 1-(1H-indol-3-yl)propan-2-amine (1.3 g, 3.73 mmol,75.4% yield, 50% purity) as yellow oil which was used in the next stepwithout further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm 7.56-7.53(m, 1H), 7.35-7.30 (m, 1H), 7.10-7.06 (m, 2H), 7.01-6.97 (m, 1H),3.21-3.17 (m, 1H), 2.85-2.79 (m, 1H), 2.73-2.69 (m, 1H), 1.15-1.10 (m,3H); ES-LCMS m/z 175.2 [M+H]⁺.

Step 3:5-(5-Fluoro-3-pyridyl)-N-[(1S)-2-(1H-indol-3-yl)-1-methyl-ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-98)

A mixture of 1-(1H-indol-3-yl)propan-2-amine (172.61 mg, 495.32 μmol,1.5 eq),7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(100 mg, 330.21 μmol, 1 eq) and DIEA (42.68 mg, 330.21 μmol, 57.52 μL, 1eq) in i-PrOH (10 mL) was degassed and purged with N₂ for 3 times, themixture was stirred at 50° C. for 3 h under N₂ atmosphere. The mixturewas concentrated under reduced pressure to give a residue which waspurified by flash silica gel chromatography (SiO₂, PE/EtOAc=10/1 to 1/1,TLC: PE/EtOAc=1/1, R_(f)=0.55) and further separated by SFC (column: OD(250 mm*30 mm, 5 μm); mobile phase: [0.1% NH₃/H₂O EtOH]; B %: 30%-30%,min) to give5-(5-fluoro-3-pyridyl)-N-[(1S)-2-(1H-indol-3-yl)-1-methyl-ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(Rt=3.870 min, 30 mg) which was purified by preparative HPLC (column:Phenomenex Synergi C18 150*30 mm*4 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 50%-80%, 12 min) to yield5-(5-fluoro-3-pyridyl)-N-[(1S)-2-(1H-indol-3-yl)-1-methyl-ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(16.33 mg, 29.93 μmol, 9.1% yield, 98.6% purity, 3HCl) ([α]²⁵_(D)=−261.788) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.67 (d,J=2.4 Hz, 1H), 8.34 (s, 1H), 8.18 (s, 1H), 7.54 (d, J=7.6 Hz, 1H), 7.45(d, J=2.0, 8.8 Hz, 1H), 7.14 (d, J=8.4 Hz, 1H), 6.99 (s, 1H), 6.95 (t,J=7.5 Hz, 1H), 6.83-6.81 (m, 1H), 5.80 (s, 1H), 4.48-4.43 (m, 1H), 3.28(d, J=2.8 Hz, 1H), 3.23-3.17 (m, 1H), 3.05-3.01 (m, 1H), 1.64 (d, J=6.4Hz, 3H), 1.34 (t, J=7.2 Hz, 6H); ES-LCMS m/z 429.3 [M+H]⁺.

Example 97

Synthesis of I-99

Synthetic Scheme:

Step 1: tert-Butyl N-(2-bromo-4,6-difluoro-phenyl)carbamate

A mixture of 2-bromo-4,6-difluoro-aniline (10 g, 48.08 mmol, 1 eq),Boc₂O (31.48 g, 144.23 mmol, 33.13 mL, 3 eq), DMAP (587.34 mg, 4.81mmol, 0.1 eq) in THE (100 mL) was degassed and purged with N₂ for 3times. The mixture was stirred at 70° C. for 16 h under N₂. The reactionmixture was concentrated under reduced pressure to give a residue whichwas dissolved in MeOH (100 mL), added K₂CO₃ (19.93 g, 144.23 mmol, 3 eq)and stirred at 70° C. for 4 h. The reaction mixture was filtered and thefiltrate was concentrated under reduced pressure. To the crude mixturewas added water (100 mL), extracted with EtOAc (100 mL×3). The combinedorganic layers were washed with brine (30 mL), dried over Na₂SO₄,filtered, concentrated under reduced pressure to give a residue whichwas purified by flash silica gel chromatography (from PE/EtOAc=1/0 to10/1, TLC: PE/EtOAc=10/1, R_(f)=0.36) to yield tert-butylN-(2-bromo-4,6-difluoro-phenyl)carbamate (12.5 g, 40.57 mmol, 84.3%yield, 100% purity) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.10(td, J=2.3, 7.7 Hz, 1H), 6.86-6.77 (m, 1H), 5.84 (s, 1H), 1.43 (s, 9H);ES-LCMS m/z 251.9, 253.9 [M-t-Bu+H]⁺.

Step 2: tert-ButylN-[2,4-difluoro-6-(2-trimethylsilylethynyl)phenyl]carbamate

A mixture of tert-butyl N-(2-bromo-4,6-difluoro-phenyl)carbamate (4 g,12.98 mmol, 1 eq), ethynyl(trimethyl)silane (1.91 g, 19.47 mmol, 2.70mL, 1.5 eq), TEA (3.94 g, 38.95 mmol, 5.42 mL, 3.0 eq), CuI (247.24 mg,1.30 mmol, 0.1 eq) and Pd(PPh₃)₂Cl₂ (455.60 mg, 649.10 μmol, 0.05 eq) inDMF (80 mL) was degassed and purged with N₂ for 3 times, then stirred at100° C. for 16 h under N₂. The reaction mixture was quenched by additionof water (300 mL), then extracted with EtOAc (200 mL×3). The combinedorganic layers were washed with brine (50 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 10/1, TLC: PE/EtOAc=20/1, R_(f)=0.31) to yieldtert-butyl N-[2,4-difluoro-6-(2-trimethylsilylethynyl)phenyl]carbamate(3.0 g, 8.20 mmol, 63.2% yield, 89.0% purity) as a black brown solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 7.00-6.93 (m, 1H), 6.90-6.81 (m, 1H), 6.09(s, 1H), 1.49 (s, 9H), 0.26 (s, 9H); ES-LCMS m/z 270.1 [M-t-Bu+H]⁺.

Step 3: 5,7-Difluoro-1H-indole

To a solution of EtOH (150 mL) was added Na (1.51 g, 65.64 mmol, 8 eq)slowly. After being stirred for 1 h at 15° C. while Na was dissolvedcompletely, to the mixture was added tert-butylN-[2,4-difluoro-6-(2-trimethylsilylethynyl)phenyl]carbamate (3.0 g, 8.20mmol, 1 eq) and the mixture was stirred at 85° C. for 16 h. The reactionmixture was concentrated under reduced pressure to remove EtOH. To theresidue was added water (100 mL), extracted with EtOAc (60 mL×3). Thecombined organic layers were washed with brine (30 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 10/1, TLC: PE/EtOAc=10/1, R_(f)=0.17) to yield5,7-difluoro-1H-indole (600 mg, 3.68 mmol, 44.8% yield, 94.0% purity) asyellow oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.36 (s, 1H), 7.27-7.25 (m,1H), 7.09 (dd, J=2.0, 9.0 Hz, 1H), 6.74 (dt, J=2.1, 10.2 Hz, 1H),6.60-6.47 (m, 1H); ES-LCMS m/z No mass was found.

Step 4: 5,7-Difluoro-1H-indole-3-carbaldehyde

To a solution of DMF (18 mL) was added POCl₃ (1.13 g, 7.37 mmol, 684.54μL, 2.0 eq) dropwise at −20° C. over a period of 10 min under N₂. After1 h, 5,7-difluoro-1H-indole (600 mg, 3.68 mmol, 1 eq) in DMF (2 mL) wasadded to the above solution during which the temperature was maintainedbelow −20° C. The reaction mixture was warmed to 15° C. and stirred at15° C. for 1 h. TLC (PE/EtOAc=1/1, R_(f)=0.40) showed the startingmaterial was consumed completely and one new spot formed. The reactionmixture was quenched by addition of NaHCO₃ (100 mL) at 0° C., extractedwith EtOAc (100 mL×3). The combined organic layers were washed withbrine (30 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by flash silicagel chromatography (from PE/EtOAc=1/0 to 1/2, TLC: PE/EtOAc=1/1,R_(f)=0.40) to yield 5,7-difluoro-1H-indole-3-carbaldehyde (500 mg, 2.35mmol, 63.7% yield, 85.0% purity) as a yellow solid. ¹H NMR (400 MHz,Acetone) δ ppm 11.72 (s, 1H), 10.05 (s, 1H), 8.36 (s, 1H), 7.73 (dd,J=2.4, 9.0 Hz, 1H), 7.02 (ddd, J=2.2, 9.4, 11.3 Hz, 1H); ES-LCMS m/z182.1 [M+H]⁺.

Step 5: 5,7-Difluoro-3-[(E)-2-nitrovinyl]-1H-indole

To a solution of 5,7-difluoro-1H-indole-3-carbaldehyde (500 mg, 2.35mmol, 1 eq) in nitromethane (15 mL) was added NH₄OAc (542.57 mg, 7.04mmol, 3.0 eq). The mixture was stirred at 110° C. for 16 h. The reactionmixture was concentrated under reduced pressure to remove nitromethane.The residue was diluted in EtOAc (50 mL), washed with water (10 mL),brine (10 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by flash silicagel chromatography (from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=1/1,R_(f)=0.70) to yield 5,7-difluoro-3-[(E)-2-nitrovinyl]-1H-indole (420mg, 1.82 mmol, 77.4% yield, 97.0% purity) as a yellow solid. ¹H NMR (400MHz, CDCl₃) δ ppm 8.23 (d, J=13.5 Hz, 1H), 7.76-7.57 (m, 2H), 7.25-7.16(m, 1H), 6.91-6.78 (m, 1H); ES-LCMS m/z 225.1 [M+H]⁺.

Step 6: 2-(5,7-Difluoro-1H-indol-3-yl)ethanamine

To a solution of 5,7-difluoro-3-[(E)-2-nitrovinyl]-1H-indole (420 mg,1.82 mmol, 1 eq) in THF (15 mL) was added dropwise LAH (1 M in THF, 6.36mL, 3.5 eq) at 0° C. After addition, the mixture was stirred at 80° C.for 2 h. TLC (PE/EtOAc=3/1, R_(f)=0.04) showed the starting material wasconsumed completely and a new spot formed. The reaction mixture wasdiluted with THF (75 mL), quenched by addition of water (0.25 mL), aq.NaOH (0.25 mL, 10% in water), water (0.75 mL) at 0° C. in sequence.After being stirred for 20 min, the mixture was filtered through celite.The filtrate was concentrated under reduced pressure to yield a crude2-(5,7-difluoro-1H-indol-3-yl)ethanamine (320 mg, 1.44 mmol, 78.9%yield, 88.0% purity) as a brown solid which was used in the next stepdirectly without further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm7.18 (s, 1H), 7.08 (dd, J=2.1, 9.4 Hz, 1H), 6.70 (ddd, J=2.3, 9.5, 11.3Hz, 1H), 2.94-2.82 (m, 4H); ES-LCMS m/z 197.1 [M+H]⁺.

Step 7:N-[2-(5,7-Difluoro-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-99)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(40 mg, 132.08 μmol, 1 eq), 2-(5,7-difluoro-1H-indol-3-yl)ethanamine(44.17 mg, 198.13 μmol, 1.5 eq) in i-PrOH (3 mL) was added DIEA (85.35mg, 660.42 μmol, 115.03 μL, 5 eq). The mixture was stirred at 60° C. for16 h. The reaction mixture was concentrated under reduced pressure toyield a residue which was purified by preparative TLC (DCM/MeOH=20/1,TLC: DCM/MeOH=20/1, R_(f)=0.48) to yieldN-[2-(5,7-difluoro-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(30.31 mg, 66.35 μmol, 50.2% yield, 98.6% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 8.82 (t, J=1.4 Hz, 1H), 8.49 (d, J=2.6 Hz,1H), 7.97-7.85 (m, 2H), 7.21-7.11 (m, 2H), 6.72-6.58 (m, 1H), 6.11 (s,1H), 3.85 (t, J=6.5 Hz, 2H), 3.30-3.24 (m, 1H), 3.15 (t, J=6.3 Hz, 2H),1.38 (d, J=7.1 Hz, 6H); ES-LCMS m/z 451.3 [M+H]⁺.

Example 98

Synthesis of I-100

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-[(4-hydroxyphenyl)methyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxamide(I-100)

To a solution of5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxylicacid (40 mg, 130.54 μmol, 1 eq) and 4-(aminomethyl)phenol (19.29 mg,156.65 μmol, 1.2 eq) in DCM (20 mL) was added HATU (74.45 mg, 195.81μmol, 1.5 eq) and DIEA (50.61 mg, 391.63 μmol, 68.21 μL, 3.0 eq). Themixture was stirred at 25° C. for 16 h. The combined reaction mixturewas diluted with H₂O (20 mL), extracted with EtOAc (20 mL×3). Thecombined organic layers were dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue which was purifiedby preparative HPLC (column: Phenomenex Synergi C18 150*30 mm*4 um;mobile phase: [water (0.05% HCl)-ACN]; B %: 47%-77%, 12 min), followedby lyophilization to yield5-(5-fluoro-3-pyridyl)-N-[(4-hydroxyphenyl)methyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidine-7-carboxamide(14.42 mg, 29.79 μmol, 22.8% yield, 98.8% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.35 (s, 1H), 8.77 (d, J=2.9 Hz,1H), 8.69 (d, J=9.3 Hz, 1H), 8.31 (s, 1H), 8.22 (s, 1H), 7.26 (d, J=8.6Hz, 2H), 6.82-6.72 (m, 2H), 4.66 (s, 2H), 3.48-3.42 (m, 1H), 1.46 (d,J=6.8 Hz, 6H); ES-LCMS m/z 406.1 [M+H]⁺.

Example 99

Synthesis of I-101

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-8-isopropyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-101)

To a solution of4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(40 mg, 137.12 umol, 1 eq) and2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethanamine (33.58 mg, 164.55 μmol, 1.2eq) in i-PrOH (10 mL) was added DIEA (53.17 mg, 411.37 μmol, 71.65 μL, 3eq). The mixture was stirred at 90° C. for 3 h. The mixture wasconcentrated under reduced pressure to give a residue which was purifiedby prep-HPLC (column: Phenomenex Synergi C18 150*30 mm*4 um; mobilephase: [water (0.05% HCl)-ACN]; B %: 20%-50%, 12 min) to yield2-(5-fluoro-3-pyridyl)-8-isopropyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(26.15 mg, 49.73 μmol, 36.27% yield, 100% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.43-9.38 (m, 1H), 9.00-8.93 (m,2H), 8.49 (d, J=5.6 Hz, 1H), 8.41 (d, J=8.2 Hz, 1H), 8.01 (s, 2H),7.59-7.54 (m, 1H), 4.19 (t, J=6.4 Hz, 2H), 3.41-3.37 (m, 2H), 3.29-3.22(m, 1H), 1.39 (d, J=6.8 Hz, 6H); ES-LCMS m/z 417.3 [M+H]⁺.

Example 100

Synthesis of I-102

Synthetic Scheme:

Step 1: 2-Methyl-4-[(E)-2-nitrovinyl]phenol

A mixture of 4-hydroxy-3-methyl-benzaldehyde (1 g, 7.34 mmol, 1 eq) andNaOAc (602.53 mg, 7.34 mmol, 1 eq) in CH₃NO₂ (10 mL) was stirred at 110°C. for 48 h. TLC (PE/EtOAc=5/1, R_(f)=0.23) indicated starting materialwas consumed completely and one new spot formed. The mixture wasconcentrated under reduced pressure to give a residue which was purifiedby flash silica gel chromatography (from PE/EtOAc=1/0 to 5/1, TLC:PE/EtOAc=5/1, R_(f)=0.23) to yield 2-methyl-4-[(E)-2-nitrovinyl]phenol(400 mg, 1.12 mmol, 15.2% yield, 50% purity) as a yellow solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 7.98 (d, J=13.6 Hz, 1H), 7.75 (d, J=13.6 Hz, 1H),7.65-7.59 (m, 1H), 7.46-7.37 (m, 1H), 6.88 (d, J=8.4 Hz, 1H), 2.24 (s,3H); ES-LCMS m/z 180.1 [M+H]⁺

Step 2: 4-(2-Aminoethyl)-2-methyl-phenol

To a solution of 2-methyl-4-[(E)-2-nitrovinyl]phenol (400 mg, 1.12 mmol,1 eq) in THE (10 mL) was added LAH (1 M, 3.35 mL, 3 eq) at 0° C. Thenthe mixture was stirred at 70° C. for 2 h. The mixture was quenched withwater (0.25 mL), 15% NaOH (0.25 mL) and water (0.75 mL). Then themixture was stirred at 20° C. for 3 h. The mixture was filtered andconcentrated under reduced pressure to yield4-(2-aminoethyl)-2-methyl-phenol (350 mg, crude) as a brown oil whichwas used in next step without further purification. ¹H NMR (400 MHz,CD₃OD) δ ppm 7.05 (s, 1H), 6.92 (s, 1H), 6.85-6.84 (m, 1H), 3.58-3.55(m, 2H), 2.17 (s, 3H), 1.60-1.57 (m, 2H); ES-LCMS m/z 152.1 [M+H]⁺.

Step 3:4-[2-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-2-methyl-phenol(I-102)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(50 mg, 165.11 μmol, 1.0 eq) and 4-(2-aminoethyl)-2-methyl-phenol (29.96mg, 198.13 μmol, 1.2 eq) in i-PrOH (10 mL) was added DIEA (64.02 mg,495.33 μmol, 86.28 μL, 3 eq). The mixture was stirred at 80° C. for 12h. The mixture was concentrated under reduced pressure to give a residuewhich was washed with MeOH (20 mL), filtered. The filtered cake waslyophilized to yield4-[2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-2-methyl-phenol(26.08 mg, 63.29 μmol, 38.3% yield, 98.4% purity) as a white solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 9.24 (s, 1H), 9.03 (s, 1H), 8.67 (s, 1H),8.40-8.36 (m, 1H), 7.99 (s, 1H), 7.92 (t, J=6.0 Hz, 1H), 7.02 (s, 1H),6.90 (dd, J=2.0, 8.2 Hz, 1H), 6.73 (s, 1H), 6.64 (d, J=8.2 Hz, 1H),3.74-3.67 (m, 2H), 3.27-3.19 (m, 1H), 2.85 (t, J=7.6 Hz, 2H), 2.07 (s,3H), 1.36 (d, J=6.8 Hz, 6H); ES-LCMS m/z 406.2 [M+H]⁺.

Example 101

Synthesis of I-103

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-103)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (30 mg, 110.76μmol, 1.0 eq) in i-PrOH (3 mL) was added DIEA (42.94 mg, 332.28 μmol,57.88 μL, 3.0 eq) and 2-(1H-indol-3-yl)ethanamine (24.00 mg, 149.80μmol, 1.35 eq). The mixture was stirred at 50° C. for 3 h. The reactionmixture was concentrated under reduced pressure to give the residuewhich was purified by preparative HPLC (HCl condition; column:Phenomenex Gemini 150×25 mm×10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 29%-49%, 10 min). The desired fraction was lyophilizedto yield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(25.41 mg, 52.74 μmol, 47.62% yield, 100% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.73 (d, J=2.5 Hz, 1H), 8.38 (s,1H), 8.26 (d, J=2.3 Hz, 1H), 7.56 (d, J=7.8 Hz, 1H), 7.46 (td, J=2.3,9.0 Hz, 1H), 7.12 (d, J=8.0 Hz, 1H), 7.00 (s, 1H), 6.96 (t, J=7.2 Hz,1H), 6.89-6.84 (m, 1H), 6.52 (d, J=2.3 Hz, 1H), 5.88 (s, 1H), 4.07 (t,J=5.9 Hz, 2H), 3.27-3.23 (m, 2H); ES-LCMS m/z 373.3 [M+H]⁺.

Example 102

Synthesis of I-104

Synthetic Scheme:

Step 1: 4-Chloro-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidine

To a solution of i-PrOH (43.10 mg, 717.06 μmol, 54.90 μL, 1 eq) in THE(8 mL) was added NaH (34.42 mg, 860.47 μmol, 60% purity, 1.2 eq). Themixture was stirred at 0° C. for 30 min.4,6-dichloro-2-(5-fluoro-3-pyridyl)pyrimidine (175 mg, 717.06 μmol, 1.0eq) was added into the above solution and the mixture was stirred at 15°C. for 12 h. The reaction mixture was concentrated under reducedpressure to remove THF. The residue was diluted with water (50 mL) andextracted with EtOAc (30 mL×3). The combined organic layers were driedover Na₂SO₄, filtered and concentrated under reduced pressure to give4-chloro-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidine (220 mg, 690.36μmol, 96.2% yield, 84.0% purity) as a yellow solid which was used in thenext step without further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm9.41 (s, 1H), 8.58 (d, J=2.8 Hz, 1H), 8.40-8.31 (m, 1H), 6.67 (s, 1H),5.53 (m, 1H), 1.43 (d, J=6.3 Hz, 6H); ES-LCMS m/z 268.1, 270.1 [M+H]⁺.

Step 2:2-(5-Fluoro-3-pyridyl)-6-isopropoxy-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrimidin-4-amine(I-104)

4-Chloro-2-(5-fluoro-3-pyridyl)-6-isopropoxy-pyrimidine (60 mg, 188.28μmol, 1.0 eq), 2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethanamine (45 mg,220.53 μmol, 1.17 eq) and DIEA (73.00 mg, 564.84 μmol, 98.38 μL, 3.0 eq)in i-PrOH (3 mL) were taken up into a microwave tube. The sealed tubewas heated at 125° C. for 6 h under microwave. The reaction mixture wasconcentrated under reduced pressure to remove solvent. The residue waspurified by preparative HPLC (HCl condition; column: Phenomenex Gemini150×25 mm×10 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 25%-55%, 10min). The desired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-6-isopropoxy-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrimidin-4-amine(20.08 mg, 38.97 μmol, 20.7% yield, 97.4% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.20 (s, 1H), 8.93 (dd, J=1.0, 2.5Hz, 1H), 8.62 (s, 1H), 8.53-8.46 (m, 2H), 8.06 (s, 1H), 7.65-7.58 (m,1H), 6.03 (s, 1H), 5.22 (s, 1H), 5.29-5.16 (m, 1H), 3.96 (m, 2H),3.30-3.27 (m, 2H), 1.42 (d, J=6.0 Hz, 6H); ES-LCMS m/z 393.2 [M+H]⁺.

Example 103

Synthesis of I-105

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-pyrrolidin-1-yl-pyrimidin-4-amine(I-105)

6-Chloro-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-4-amine(80 mg, 215.33 μmol, 1.0 eq), pyrrolidine (1.28 g, 17.97 mmol, 1.5 mL,83.45 eq) and DIEA (139.15 mg, 1.08 mmol, 187.53 μL, 5.0 eq) were takenup into a microwave tube in i-PrOH (1.5 mL). The sealed tube was heatedat 130° C. for 3 h under microwave. The reaction mixture wasconcentrated under reduced pressure to remove solvent. The residue waspurified by preparative HPLC (HCl condition; column: Phenomenex Gemini150×25 mm×10 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 25%-55%, 10min). The desired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-pyrrolidin-1-yl-pyrimidin-4-amine(47.28 mg, 91.26 μmol, 42.3% yield, 98.8% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.00 (s, 1H), 8.77 (s, 1H), 8.22(s, 1H), 7.64 (d, J=7.3 Hz, 1H), 7.33 (d, J=7.9 Hz, 1H), 7.16-6.98 (m,4H), 3.72 (d, J=6.0 Hz, 4H), 3.15 (t, J=5.8 Hz, 4H), 2.00 (s, 4H);ES-LCMS m/z 403.3 [M+H]⁺.

Example 104

Synthesis of I-106

Synthetic Scheme:

Step 1: 1-Amino-3-methyl-butan-2-ol

A mixture of 2-isopropyloxirane (600 mg, 6.97 mmol, 1 eq) in NH₃.H₂O (5mL) was degassed and purged with N₂ for 3 times. Then the mixture wasstirred at 10-15° C. for 16 h under N₂ atmosphere. The reaction mixturewas concentrated under reduced pressure to give the crude product1-amino-3-methyl-butan-2-ol (700 mg, 6.79 mmol, 97.4% yield, crude) ascolorless oil which was used in the next step without furtherpurification. ¹H NMR (400 MHz, CDCl₃) δ ppm 3.39-3.22 (m, 1H), 2.91-2.72(m, 1H), 2.60-2.49 (m, 1H), 1.71-1.62 (m, 1H), 0.98-0.91 (m, 6H);ES-LCMS m/z No correct mass was found.

Step 2: 1-[(3-Chloropyrazin-2-yl)amino]-3-methyl-butan-2-ol

A mixture of 2,3-dichloropyrazine (670 mg, 4.50 mmol, 1 eq) and1-amino-3-methyl-butan-2-ol (695.93 mg, 6.75 mmol, 1.5 eq) in1,4-dioxane (3 mL) was degassed and purged with N₂ for 3 times. Themixture was stirred at 110° C. for 16 h under N₂ atmosphere. TLC(PE/EtOAc=5/1, R_(f)=0.54) showed the starting material was not consumedcompletely and a new spots was found. The reaction mixture wasconcentrated under reduced pressure to give a residue which was purifiedon silica gel column chromatography (from PE/EtOAc=1/0 to 5/1, TLC:PE/EtOAc=5/1, R_(f)=0.54) to give the product1-[(3-chloropyrazin-2-yl)amino]-3-methyl-butan-2-ol (500 mg, 2.27 mmol,50.5% yield, 98.0% purity) as colorless oil. ¹H NMR (400 MHz, CDCl₃) δppm 7.92 (d, J=2.9 Hz, 1H), 7.60 (d, J=2.6 Hz, 1H), 5.61 (br s, 1H),3.74 (ddd, J=2.4, 6.5, 13.8 Hz, 1H), 3.59-3.58 (m, 1H), 3.44-3.37 (m,1H), 2.83 (d, J=4.4 Hz, 1H), 1.89-1.72 (m, 1H), 1.03 (dd, J=6.8, 10.6Hz, 6H); ES-LCMS m/z 216.0, 218.1 [M+H]⁺.

Step 3: 1-[(3,5-Dichloropyrazin-2-yl)amino]-3-methyl-butan-2-ol

A mixture of 1-[(3-chloropyrazin-2-yl)amino]-3-methyl-butan-2-ol (380mg, 1.73 mmol, 1 eq) and NCS (276.68 mg, 2.07 mmol, 1.2 eq) in CHCl₃ (10mL) was degassed and purged with N₂ for 3 times and stirred at 70° C.for 2 h under N₂ atmosphere. The reaction mixture was concentrated underreduced pressure to give a residue which was purified on silica gelcolumn chromatography (from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1,R_(f)=0.46) to give the product of1-[(3,5-dichloropyrazin-2-yl)amino]-3-methyl-butan-2-ol (345 mg, 1.23mmol, 71.1% yield, 89.0% purity) as yellow oil. ¹H NMR (400 MHz, CDCl₃)δ ppm 7.95 (s, 1H), 5.59 (br s, 1H), 3.74 (ddd, J=2.8, 6.7, 13.7 Hz,1H), 3.58-3.56 (m, 1H), 3.38-3.33 (m, 1H), 1.83-1.73 (m, 1H), 1.03 (dd,J=6.8, 10.1 Hz, 6H); ES-LCMS m/z 250.0, 252.0 [M+H]⁺.

Step 4: 1-[(3,5-Dichloropyrazin-2-yl)amino]-3-methyl-butan-2-one

To a solution of 1-[(3,5-dichloropyrazin-2-yl)amino]-3-methyl-butan-2-ol(405 mg, 1.44 mmol, 1 eq) in DCM (30 mL) was added Dess-Martin (733.47mg, 1.73 mmol, 1.2 eq). The mixture was stirred at 20° C. for 2 h. TLC(PE/EtOAc=3/1, R_(f)=0.51) showed about 50% of starting material wasremained. Dess-Martin (300 mg, 0.71 mmol) was added and the reaction wasstirred at 20° C. for 16 h. TLC (PE/EtOAc=3/1, R_(f)=0.51) showed about20% of starting material was remained. The reaction mixture was quenchedby addition sat Na₂S₂O₃ solution (50 mL), extracted with EtOAc (50mL×3). The combined organic layers were dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a residue whichwas purified on silica gel column chromatography (from PE/EtOAc=1/0 to3/1, TLC: PE/EtOAc=3/1, R_(f)=0.51) to give the product of1-[(3,5-dichloropyrazin-2-yl)amino]-3-methyl-butan-2-one (200 mg, 669.06μmol, 46.4% yield, 83.0% purity) as a yellow oil. ¹H NMR (400 MHz,CDCl₃) δ ppm 7.96 (s, 1H), 6.03 (br s, 1H), 4.35 (d, J=4.4 Hz, 2H), 2.76(m, 1H), 1.22 (d, J=6.8 Hz, 6H); ES-LCMS m/z 247.9, 249.9 [M+H]⁺.

Step 5: 6,8-Dichloro-3-isopropyl-imidazo[1,2-a]pyrazine

To a solution of1-[(3,5-dichloropyrazin-2-yl)amino]-3-methyl-butan-2-one (180 mg, 602.16μmol, 1 eq) in TFA (2 mL) was added TFAA (379.41 mg, 1.81 mmol, 251.27μL, 3 eq). The mixture was stirred at 20° C. for 5 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified on silica gel column chromatography (from PE/EtOAc=1/0 to10/3, TLC: PE/EtOAc=3/1, R_(f)=0.48) to give the product6,8-dichloro-3-isopropyl-imidazo[1,2-a]pyrazine (120 mg, 521.53 μmol,86.6% yield, 100.0% purity) as a light yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 7.94 (s, 1H), 7.69 (s, 1H), 3.22-3.16 (m, 1H), 1.44 (d,J=6.8 Hz, 6H); ES-LCMS m/z 230.0, 232.0 [M+H]⁺.

Step 6:6-Chloro-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-imidazo[1,2-a]pyrazin-8-amine

To a solution of 6,8-dichloro-3-isopropyl-imidazo[1,2-a]pyrazine (50 mg,217.30 μmol, 1 eq) and 2-(1H-indol-3-yl)ethanamine (41.78 mg, 260.76μmol, 1.2 eq) in i-PrOH (3 mL) was added DIEA (84.25 mg, 651.91 μmol,113.55 μL, 3.0 eq). The mixture was stirred at 45° C. for 3 h. Thereaction mixture was concentrated under reduced pressure to give aresidue which was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=1/1, R_(f)=0.44) to give the product6-chloro-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-imidazo[1,2-a]pyrazin-8-amine(60 mg, 162.78 μmol, 74.9% yield, 96.0% purity) as brown oil. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.07 (br s, 1H), 7.71 (d, J=7.9 Hz, 1H), 7.38 (d,J=7.9 Hz, 1H), 7.30 (s, 1H), 7.24-7.19 (m, 2H), 7.17-7.12 (m, 1H), 7.11(d, J=2.2 Hz, 1H), 6.21 (br s, 1H), 4.02-3.89 (m, 2H), 3.17 (t, J 6.8Hz, 2H), 3.12-3.02 (m, 1H), 1.37 (d, J=7.1 Hz, 6H); ES-LCMS m/z 354.0,356.1 [M+H]⁺.

Step 7:6-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-imidazo[1,2-a]pyrazin-8-amine(I-106)

6-Chloro-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-imidazo[1,2-a]pyrazin-8-amine(60 mg, 162.78 μmol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (45.87 mg,325.56 μmol, 2.0 eq), Pd(dppf)Cl₂ (11.91 mg, 16.28 μmol, 0.1 eq) andCs₂CO₃ (159.11 mg, 488.35 μmol, 3.0 eq) were taken up into a microwavetube in 1,4-dioxane (2 mL) and H₂O (0.5 mL). The mixture was purged withN₂ for 3 min. The sealed tube was heated at 110° C. for 30 min undermicrowave. The reaction mixture was diluted with (20 mL) and extractedwith ethyl acetate (20 mL×3). The combined organic layers were driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto give a residue which was purified by preparative HPLC (column:Phenomenex Synergi C18 150*30 mm*4 um; mobile phase: [water(0.05%HCl)-ACN]; B %: 30%-60%, 12 min), followed by lyophilization to yield6-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-imidazo[1,2-a]pyrazin-8-amine(50.13 mg, 94.00 μmol, 57.8% yield, 98.2% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.21 (br s, 1H), 8.86 (br s, 1H),8.74 (d, J=9.0 Hz, 1H), 8.57 (s, 1H), 7.89 (s, 1H), 7.51 (d, J=7.8 Hz,1H), 7.18 (d, J=8.3 Hz, 1H), 7.09 (s, 1H), 6.94 (t, J=7.5 Hz, 1H),6.89-6.81 (m, 1H), 4.06 (t, J=6.1 Hz, 2H), 3.46 (d, J=6.0 Hz, 1H), 3.18(t, J=6.3 Hz, 2H), 1.42 (d, J=6.3 Hz, 6H); ES-LCMS m/z 415.3 [M+H]⁺.

Example 105

Synthesis of I-107

Synthetic Scheme:

Step 1:N-[2-(5,7-Difluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-107)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(120 mg, 404.51 μmol, 1 eq) and2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethanamine (182.19 mg, 606.76μmol, 1.5 eq, oxalic acid) in i-PrOH (20 mL) was added DIEA (261.40 mg,2.02 mmol, 352.29 μL, 5.0 eq). The mixture was stirred at 70° C. for 18h. The reaction mixture concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: PhenomenexGemini 150*25 mm*10 um; mobile phase: [water (0.05% ammonia hydroxidev/v)-ACN]; B %: 63%-93%, 10 min), followed by lyophilization to yieldN-[2-(5,7-difluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(94.41 mg, 196.14 μmol, 48.5% yield, 96.5% purity, 92.85 mg was fordelivery) as a light yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.16(s, 1H), 8.96 (s, 1H), 8.61 (d, J=2.8 Hz, 1H), 8.06-7.95 (m, 3H), 7.20(dd, J=2.0, 9.5 Hz, 1H), 6.80-6.68 (m, 1H), 6.34 (s, 1H), 3.69 (q, J=6.5Hz, 2H), 3.19 (spt, J=6.8 Hz, 1H), 2.99 (t, J=6.7 Hz, 2H), 2.16 (s, 3H),1.33 (d, J=7.0 Hz, 6H); ES-LCMS m/z 465.3 [M+H]⁺.

Example 106

Synthesis of I-108

Synthetic Scheme:

Step 1:4-[2-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenol (I-108)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(52.08 mg, 171.99 umol, 1 eq) in i-PrOH (3 mL) was added4-(2-aminoethyl)phenol (28.31 mg, 206.38 μmol, 1.2 eq) and DIEA (66.68mg, 515.96 μmol, 89.87 μL, 3 eq). The mixture was stirred at 60° C. for16 h. The mixture was concentrated to afford the crude product which waspurified by preparative HPLC (MeCN/H₂O as eluents, acidic condition,Instrument: Phenomenex Gemini 150*25 mm*10 μm/Mobile phase: water(0.05%HCl)-CAN/Gradient: B from 42% to 72% in 10 min/Flow rate: 25 mL/min),followed by lyophilization to yield4-[2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenol (49.37 mg, 105.68 μmol, 61.5% yield, 99.4% purity,2HCl) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.83 (d, J=2.5Hz, 2H), 8.27 (s, 1H), 8.16 (dd, J=1.8, 8.8 Hz, 1H), 7.08 (d, J=8.3 Hz,2H), 6.63 (d, J=8.3 Hz, 2H), 6.38 (s, 1H), 3.97 (t, J=6.5 Hz, 2H),3.38-3.34 (m, 1H), 3.01 (t, J=6.5 Hz, 2H), 1.39 (d, J=7.0 Hz, 6H);ES-LCMS m/z 392.1 [M+H]⁺.

Example 107

Synthesis of I-109

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-(5-methoxyindan-2-yl)pyrazolo[1,5-a]pyrimidin-7-amine

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(80 mg, 264.17 μmol, 1.0 eq) in i-PrOH (3 mL) was added DIEA (102.42 mg,792.51 μmol, 138.04 μL, 3.0 eq) and 5-methoxyindan-2-amine (60 mg,367.61 μmol, 1.39 eq). The mixture was stirred at 50° C. for 15 h. Thereaction mixture was concentrated under reduced pressure to give aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=100/1 to 2/1, TLC: PE/EtOAc=3/1, R_(f)=0.70) to yield5-(5-fluoro-3-pyridyl)-3-isopropyl-N-(5-methoxyindan-2-yl)pyrazolo[1,5-a]pyrimidin-7-amine(120 mg, 155.51 μmol, 58.9% yield, 54.1% purity) as a yellow oil. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.08 (s, 1H), 8.55 (d, J=2.8 Hz, 1H), 8.21 (td,J=2.3, 9.5 Hz, 1H), 7.87 (s, 1H), 7.18 (d, J=8.3 Hz, 1H), 6.83 (s, 1H),6.57 (d, J=7.5 Hz, 1H), 6.41 (s, 1H), 4.70-4.60 (m, 1H), 3.81 (s, 3H),3.50 (ddd, J=6.8, 12.6, 15.7 Hz, 2H), 3.42-3.30 (m, 1H), 3.08 (dt,J=4.5, 16.1 Hz, 2H), 1.42 (d, J=6.8 Hz, 6H); ES-LCMS m/z 418.1 [M+H]⁺.

Step 2:2-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(I-109)

To a solution of5-(5-fluoro-3-pyridyl)-3-isopropyl-N-(5-methoxyindan-2-yl)pyrazolo[1,5-a]pyrimidin-7-amine(95 mg, 123.11 μmol, 1 eq) in HBr (9 mL) was stirred at 120° C. for 2 h.The reaction mixture was concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (HCl condition; column:Phenomenex Gemini 150×25 mm×10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 49%-79%, 10 min). The desired fraction was lyophilizedto yield2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol (30.69 mg, 61.40 μmol, 49.8% yield, 95.3% purity, 2HClsalt) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.00 (s, 1H),8.83 (d, J=2.5 Hz, 1H), 8.35 (td, J=2.3, 9.0 Hz, 1H), 8.24 (s, 1H), 7.08(d, J=8.3 Hz, 1H), 6.96 (s, 1H), 6.71 (s, 1H), 6.65 (dd, J=2.3, 8.0 Hz,1H), 5.02 (t, J=6.9 Hz, 1H), 3.51-3.40 (m, 2H), 3.40-3.32 (m, 1H),3.25-3.12 (m, 2H), 1.39 (d, J=7.0 Hz, 6H); ES-LCMS m/z 404.2 [M+H]⁺.

Example 108

Synthesis of I-110

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-110)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(45 mg, 123.83 umol, 1.0 eq) in i-PrOH (2 mL) was added DIEA (48.01 mg,371.49 μmol, 64.70 μL, 3.0 eq) and2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethanamine (25 mg, 155.08 μmol, 1.25eq). The mixture was stirred at 50° C. for 15 h. The reaction mixturewas concentrated under reduced pressure to give the residue which waspurified by preparative HPLC (HCl condition; column: Phenomenex Gemini150×25 mm×10 um; mobile phase: [water(0.05% HCl)-ACN]; B %: 20%-50%, 10min). The desired fraction was lyophilized to yield5-(5-fluoro-3-pyridyl)-3-isopropyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(16.32 mg, 30.38 μmol, 24.5% yield, 97.7% purity, 3HCl salt) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.97 (s, 1H), 8.84 (d, J=2.4 Hz,1H), 8.57-8.52 (m, 2H), 8.35 (td, J=2.1, 9.0 Hz, 1H), 8.23 (s, 1H), 8.10(s, 1H), 7.70-7.65 (m, 1H), 6.90 (s, 1H), 4.15 (t, J=6.9 Hz, 2H), 3.45(t, J=6.9 Hz, 2H), 3.38-3.33 (m, 1H), 1.38 (d, J=6.8 Hz, 6H); ES-LCMSm/z 416.2 [M+H]⁺.

Example 109

Synthesis of I-120

Synthetic Scheme:

Step 1: 4,6-Dichloro-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine

To a mixture of 2-(1H-indol-3-yl)ethanamine (2.96 g, 18.50 mmol, 1.05eq) in THF (50 mL) was added NaH (1.06 g, 26.42 mmol, 1.5 eq). Themixture was stirred for 30 min at 0° C. under N₂ atmosphere. A solutionof 4,6-dichloro-2-methylsulfonyl-pyrimidine (4 g, 17.62 mmol, 1 eq) inTHF (50 mL) was added to the mixture dropwise. The mixture was stirredat −55° C. for 12 h. To the reaction mixture was added 1N NH₄Cl solution(5 mL) and concentrated. The residue was purified by flash silica gelchromatography (PE/EtOAc=3/1, R_(f)=0.5) to yield4,6-dichloro-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine (2.1 g, 5.81mmol, 33.0% yield, 85% purity) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 8.68 (s, 1H), 8.00 (d, J=10.8 Hz, 1H), 7.64 (t, J=6.1 Hz,1H), 7.39-7.33 (m, 1H), 7.26-7.20 (m, 1H), 7.16-7.09 (m, 1H), 7.08-6.99(m, 1H), 6.61-6.53 (m, 1H), 3.84-3.70 (m, 2H), 3.05 (d, J=6.1 Hz, 2H);ES-LCMS m/z 307.0, 309.1 [M+H]⁺.

Step 2:4-Chloro-6-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine

To a mixture of 4,6-dichloro-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine(300 mg, 830.14 mol, 1 eq) and (5-fluoro-3-pyridyl)boronic acid (93.58mg, 664.11 mol, 0.8 eq) in 1,4-dioxane (3 mL) and H₂O (2 mL) was addedCs₂CO₃ (540.95 mg, 1.66 mmol, 2 eq) and Pd(dppf)Cl₂ (60.74 mg, 83.01mol, 0.1 eq) under N₂ atmosphere. The mixture was stirred at 110° C. for20 min under microwave. The reaction mixture was concentrated. Theresidue was purified on silica gel column chromatography (from pure PEto PE/EtOAc=3/1, TLC: PE/EtOAc=3/1, R_(f)=0.33) to yield4-chloro-6-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine(70 mg, 171.29 mol, 20.6% yield, 90% purity) as a light yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.91 (br s, 1H), 8.50 (br s, 1H), 7.97 (br s,2H), 7.60 (d, J=7.1 Hz, 1H), 7.32 (d, J=8.1 Hz, 1H), 7.15 (t, J=7.6 Hz,1H), 7.10-7.04 (m, 1H), 7.02 (br s, 1H), 6.92 (s, 1H), 5.39 (br s, 1H),3.79 (br s, 2H), 3.05 (t, J=6.7 Hz, 2H); ES-LCMS m/z 368.1, 370.1[M+H]⁺.

Step 3:4-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine(I-120)

To a solution of4-chloro-6-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine(70 mg, 171.29 mol, 1 eq) in MeOH (5 mL) and NH₃—H₂O (0.2 mL) was addedPd/C (30 mg, 10% purity). The suspension was degassed under vacuum andpurged with H₂ several times. The mixture was stirred under H₂ (15 psi)atmosphere at 10° C. for 0.5 h. The reaction mixture was filtered andconcentrated. The residue was purified by preparative HPLC (column:Agela ASB 150*25 mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %:33%-63%, 8 min), followed by lyophilization to yield4-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine (20.8mg, 50.20 □mol, 29.3% yield, 98.1% purity, 2 HCl) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 9.06 (br s, 1H), 8.74 (br s, 1H), 8.32-8.06(m, 2H), 7.59-7.55 (m, 1H), 7.43 (d, J=6.6 Hz, 1H), 7.24-7.04 (m, 2H),7.01-6.93 (m, 2H), 4.05-3.96 (m, 2H), 3.15 (t, J=6.4 Hz, 2H); ES-LCMSm/z 334.1 [M+H]⁺.

Example 110

Synthesis of I-121

Synthetic Scheme:

Step 1(3R)—N-(2-Methylsulfanylpyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine

To a solution of4-chloro-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazine (500 mg, 1.69mmol, 1 eq, HCl) in MeCN (30 mL) was added DIEA (1.31 g, 10.12 mmol,1.76 mL, 6 eq) and (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (314.20mg, 1.69 mmol, 1 eq). The mixture was stirred at 50° C. for 2 h. Themixture was concentrated. The crude material was purified on silica gelcolumn chromatography (from pure PE to PE/EtOAc=2/1, TLC: PE/EtOAc=3/1,R_(f)=0.4) to yield(3R)—N-(2-methylsulfanylpyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(300 mg, 770.46 μmol, 45.7% yield, 90% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 7.90-7.77 (m, 2H), 7.44 (d, J=7.8 Hz, 1H), 7.30(d, J=8.1 Hz, 1H), 7.18-7.12 (m, 1H), 7.11-7.06 (m, 1H), 6.61 (d, J=8.3Hz, 1H), 6.21 (s, 1H), 3.28 (dd, J=5.0, 15.5 Hz, 1H), 3.00-2.78 (m, 4H),2.55 (s, 3H), 2.31-2.15 (m, 2H); ES-LCMS m/z 351.2 [M+H]⁺.

Step 2: tert-Butyl(3R)-3-[tert-butoxycarbonyl-(2-methylsulfanylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

To a solution of(3R)—N-(2-methylsulfanylpyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(200 mg, 513.64 μmol, 1 eq) in 1,4-dioxane (30 mL) was added DMAP(188.25 mg, 1.54 mmol, 3 eq) and Boc₂O (672.60 mg, 3.08 mmol, 708.00 μL,6 eq). The mixture was stirred at 110° C. for 24 h. The mixture wasconcentrated. The crude material was purified on silica gel columnchromatography (from pure PE to PE/EtOAc=3/1, TLC: PE/EtOAc=5/1,R_(f)=0.5) to yield tert-butyl(3R)-3-[tert-butoxycarbonyl-(2-methylsulfanylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(300 mg, 492.49 μmol, 95.9% yield, 90.4% purity) as an off white solid.¹H NMR (400 MHz, CDCl₃) δ ppm 8.11-8.02 (m, 2H), 7.33 (d, J=7.6 Hz, 1H),7.23-7.14 (m, 2H), 6.41 (s, 1H), 4.79-4.72 (m, 1H), 3.31-3.01 (m, 4H),2.56 (s, 3H), 2.41-2.17 (m, 2H), 1.64 (s, 9H), 1.32 (s, 9H); ES-LCMS m/z551.3 [M+H]⁺.

Step 3: tert-Butyl(3R)-3-[(8-bromo-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

To a solution of tert-butyl(3R)-3-[tert-butoxycarbonyl-(2-methylsulfanylpyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(260 mg, 426.82 μmol, 1 eq) in MeCN (20 mL) and DCM (20 mL) was addedNBS (79.76 mg, 448.17 μmol, 1.05 eq). The mixture was stirred at 10° C.for 30 min. The mixture was concentrated. The residue was purified onsilica gel column chromatography (from PE/EtOAc=5/1 to 2/1, TLC:PE/EtOAc=5/1, R_(f)=0.58) to yield tert-butyl(3R)-3-[(8-bromo-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(200 mg, 299.89 μmol, 70.3% yield, 94.4% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.09 (d, J=7.8 Hz, 1H), 8.05 (s, 1H), 7.34(d, J=8.0 Hz, 1H), 7.25-7.17 (m, 2H), 4.83-4.73 (m, 1H), 3.34-3.25 (m,1H), 3.23-3.02 (m, 3H), 2.62 (s, 3H), 2.36 (td, J=6.2, 12.0 Hz, 1H),2.23-2.21 (m, 1H), 1.66 (s, 9H), 1.36 (s, 9H); ES-LCMS m/z 629.2, 631.2[M+H]⁺.

Step 4: tert-Butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropenyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

To a solution of tert-butyl(3R)-3-[(8-bromo-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(200 mg, 299.89 μmol, 1 eq),2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (75.59 mg, 449.83μmol, 1.5 eq) in 1,4-dioxane (4 mL) and H₂O (2 mL) was added Cs₂CO₃(293.13 mg, 899.67 μmol, 3 eq) and Pd(dppf)Cl₂ (21.94 mg, 29.99 μmol,0.1 eq) under N₂ atmosphere. The mixture was stirred at 110° C. for 1 hunder microwave. After filtration, the filtrate was concentrated. Thecrude material was purified on silica gel column chromatography (frompure PE to PE/EtOAc=5/1, TLC: PE/EtOAc=5/1, R_(f)=0.65) to yieldtert-butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropenyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(180 mg, 274.23 μmol, 91.5% yield, 90% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.13-8.05 (m, 2H), 7.32 (d, J=7.6 Hz, 1H),7.23-7.15 (m, 2H), 5.86 (s, 1H), 5.12 (s, 1H), 4.79-4.74 (m, 1H),3.29-3.01 (m, 4H), 2.57 (s, 3H), 2.33-2.18 (m, 5H), 1.34 (s, 9H), 1.26(s, 9H); ES-LCMS m/z 591.3 [M+H]⁺.

Step 5: tert-Butyl(3R)-3-[tert-butoxycarbonyl-[2-(3-fluorophenyl)-8-isopropenyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

A mixture of tert-butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropenyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(140 mg, 213.29 μmol, 1 eq), (3-fluorophenyl)boronic acid (89.53 mg,639.88 μmol, 3 eq), Pd(PPh₃)₄ (24.65 mg, 21.33 umol, 0.1 eq) and CuTC(122.02 mg, 639.88 μmol, 3 eq) in THE (2 mL) was taken up into amicrowave tube under N₂ atmosphere. The mixture was stirred at 100° C.for 3 h under microwave. After filtration, the filtrate wasconcentrated. The residue was purified with preparative TLC(PE/EtOAc=5/1, R_(f)=0.68) to afford tert-butyl(3R)-3-[tert-butoxycarbonyl-[2-(3-fluorophenyl)-8-isopropenyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(70 mg, 64.11 μmol, 30.1% yield, 58.5% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.21-8.13 (m, 2H), 8.02 (d, J=8.1 Hz, 2H),7.31-7.23 (m, 2H), 7.13-7.06 (m, 3H), 5.97 (s, 1H), 5.17 (s, 1H),4.85-4.83 (m, 1H), 3.10-2.97 (m, 4H), 2.22-2.14 (m, 5H), 1.29 (d, J=4.4Hz, 18H); ES-LCMS m/z 639.4 [M+H]⁺.

Step 6:(3R)—N-[2-(3-Fluorophenyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-121)

To a solution of tert-butyl(3R)-3-[tert-butoxycarbonyl-[2-(3-fluorophenyl)-8-isopropenyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(60 mg, 54.95 μmol, 1 eq) and NH₃.H₂O (546.00 mg, 4.36 mmol, 600.00 μL,28% purity, 79.38 eq) in MeOH (15 mL) was added Pd/C (30 mg, 9.16 umol,10% purity). The mixture was stirred under H₂ atmosphere (15 psi) at 25°C. for 6 h. After filtration, the filtrate was concentrated. To theresidue was added DCM (3 mL) and TFA (1.54 g, 13.51 mmol, 1 mL, 245.78eq). The mixture was stirred at 15° C. for 1 h. The mixture wasconcentrated to yield a residue which was purified by preparative HPLC(column: Agela ASB 150*25 mm*5 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 80%-100%, 8 min), followed by lyophilization to yield(3R)—N-[2-(3-fluorophenyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(12.51 mg, 26.23 μmol, 9.8% yield, 100% purity, HCl, OR: [α]^(22.4)_(D)=+9.972, (MeOH, c=0.071 g/100 mL)) as a yellow solid. ¹H NMR (400MHz, CD₃OD) δ ppm 8.24 (d, J=8.1 Hz, 1H), 8.09 (d, J=10.3 Hz, 1H), 7.95(s, 1H), 7.45 (dt, J=5.9, 7.9 Hz, 1H), 7.37 (d, J=7.8 Hz, 1H), 7.26 (d,J=7.8 Hz, 1H), 7.20 (dt, J=2.1, 8.3 Hz, 1H), 7.02 (t, J=7.2 Hz, 1H),6.97-6.91 (m, 1H), 4.89-4.85 (m, 1H), 3.28-3.23 (m, 1H), 3.08-2.87 (m,3H), 2.43-2.33 (m, 1H), 2.30-2.19 (m, 1H), 1.39 (d, J=6.8 Hz, 6H);ES-LCMS m/z 441.3 [M+H]⁺.

Example 111

Synthesis of I-122

Synthetic Scheme:

Step 1: Ethyl N-(1H-pyrazol-5-ylcarbamothioyl)carbamate

To a suspension of 3H-pyrazol-3-amine (15.84 g, 190.62 mmol, 1 eq) inDCM (160 mL) was added ethoxycarbonyl isothiocyanate (25 g, 190.62 mmol,22.52 mL, 1 eq) at 0° C. The mixture was stirred at 25° C. for 12 hunder N₂ atmosphere. The mixture was filtered. The filter cake waswashed with DCM (20 mL×2), dried to yield ethylN-(1H-pyrazol-5-ylcarbamothioyl)carbamate (26 g, 109.22 mmol, 57.3%yield, 90% purity) as a white solid. ¹H NMR (400 MHz, CD₃OD) S ppm 7.59(d, J=2.2 Hz, 1H), 7.12 (s, 1H), 4.27 (q, J=7.2 Hz, 2H), 1.34 (t, J=7.1Hz, 3H); ES-LCMS m/z 215.1 [M+H]⁺.

Step 2: 2-Thioxo-1H-pyrazolo[1,5-a][1,3,5]triazin-4-one

To a mixture of ethyl N-(1H-pyrazol-5-ylcarbamothioyl)carbamate (25.5 g,107.12 mmol, 1 eq) in MeCN (300 mL) was added K₂CO₃ (44.42 g, 321.36mmol, 3 eq). The mixture was stirred at 85° C. for 4 h. The mixturediluted with water (100 mL) and adjusted pH to 5-6 with 2 N HCl. Thesolvents were evaporated and the residue was suspended in water (600mL). The solid was filtered off, washed with water (60 mL×2), dried toyield 2-thioxo-1H-pyrazolo[1,5-a][1,3,5]triazin-4-one (16 g, 90.38 mmol,84.4% yield, 95% purity) as a white solid. ¹H NMR (400 MHz, CD₃OD) S ppm7.83 (s, 1H), 5.93 (d, J=1.7 Hz, 1H); ES-LCMS m/z 169.1 [M+H]⁺.

Step 3: 2-Methylsulfanyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

To a suspension of 2-thioxo-1H-pyrazolo[1,5-a][1,3,5]triazin-4-one (16g, 90.38 mmol, 1 eq) in EtOH (120 mL) was added NaOH (7.23 g, 180.76mmol, 2 eq) in H₂O (9.6 mL), MeI (15.96 g, 112.44 mmol, 7.0 mL, 1.24 eq)was added dropwise. The mixture was stirred at 15° C. for 1 h. Themixture was acidified with 1N aq. HCl (80 mL) and EtOH was evaporated.The mixture was filtered, washed with H₂O (50 mL×2). The filter cake wasdried to yield 2-methylsulfanyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one(16 g, 83.42 mmol, 92.3% yield, 95% purity) as a white solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 7.74 (d, J=2.0 Hz, 1H), 5.99 (d, J=2.2 Hz, 1H),2.48 (s, 3H); ES-LCMS m/z 183.2 [M+H]⁺.

Step 4: 8-Bromo-2-methylsulfanyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-on

To a solution of 2-methylsulfanyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one(3.0 g, 15.64 mmol, 1 eq) in DMF (30 mL) was added NBS (2.51 g, 14.08mmol, 0.9 eq). The mixture was stirred at 15° C. for 0.5 h. The mixturewas concentrated to yield8-bromo-2-methylsulfanyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (4 g,12.26 mmol, 78.4% yield, 80% purity) as green oil. ¹H NMR (400 MHz,CD₃OD) S ppm 7.72 (s, 1H), 2.49 (s, 3H); ES-LCMS m/z 261.0, 263.0[M+H]⁺.

Step 5: 8-Bromo-4-chloro-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazine

A suspension of8-bromo-2-methylsulfanyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (2.0 g,6.13 mmol, 1 eq) and N,N-dimethylaniline (742.58 mg, 6.13 mmol, 776.76μL, 1 eq) in POCl₃ (16.50 g, 107.61 mmol, 10 mL, 17.56 eq) was stirredat 130° C. for 4 h. The mixture was added into water (100 mL) at 0° C.,extracted with EtOAc (200 mL×3). The combined organic layers were washedwith brine (50 mL), dried over Na₂SO₄, filtered and concentrated. Thecrude material was purified on silica gel column chromatography (frompure PE to PE/EtOAc=3/1, TLC: PE/EtOAc=3/1, R_(f)=0.8) to yield8-bromo-4-chloro-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazine (600mg, 2.08 mmol, 33.9% yield, 97% purity) as a white solid. ¹H NMR (400MHz, CDCl₃) δ ppm 8.12 (s, 1H), 2.66 (s, 3H); ES-LCMS m/z 278.9, 280.9[M+H]⁺.

Step 6:(3R)—N-(8-Bromo-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine

To a solution of8-bromo-4-chloro-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazine (600mg, 2.08 mmol, 1 eq) in i-PrOH (12 mL) was added DIEA (807.22 mg, 6.25mmol, 1.09 mL, 3 eq) and (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(426.55 mg, 2.29 mmol, 1.10 eq). The mixture was stirred at 50° C. for 2h. The mixture was concentrated, extracted with EtOAc (60 mL×3). Thecombined organic layers were washed with brine (50 mL), dried overNa₂SO₄, filtered and concentrated to yield(3R)—N-(8-bromo-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(800 mg, 1.68 mmol, 80.6% yield, 90% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 7.86-7.80 (m, 2H), 7.45 (d, J=7.6 Hz, 1H), 7.32(d, J=8.1 Hz, 1H), 7.17 (t, J=6.8 Hz, 1H), 7.13-7.07 (m, 1H), 6.59 (d,J=8.3 Hz, 1H), 4.82-4.78 (m, 1H), 3.31-3.26 (m, 1H), 2.99-2.84 (m, 3H),2.61 (s, 3H), 2.30-2.20 (m, 2H); ES-LCMS m/z 429.1, 431.1 [M+H]⁺.

Step 7:(3R)—N-(8-Isopropenyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine

To a mixture of(3R)—N-(8-bromo-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(400 mg, 838.50 μmol, 1 eq), Pd(dppf)Cl₂ (61.35 mg, 83.85 μmol, 0.1 eq),Cs₂CO₃ (683.00 mg, 2.10 mmol, 2.5 eq) in 1,4-dioxane (10 mL) and H₂O(2.5 mL) was added 2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(704.51 mg, 4.19 mmol, 5 eq) under N₂ atmosphere. The mixture wasstirred at 110° C. for 1.5 h under N₂ atmosphere under microwave. Thecombined reaction mixture was concentrated, extracted with EtOAc (50mL×3). The combined organic layers were washed with brine (50 mL), driedover Na₂SO₄, filtered and concentrated. The crude material was purifiedon silica gel column chromatography (from pure PE to PE/EtOAc=3/1, TLC:PE/EtOAc=3/1, R_(f)=0.46) to yield(3R)—N-(8-isopropenyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(250 mg, 550.57 μmol, 65.6% yield, 86% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 7.87 (s, 1H), 7.81 (s, 1H), 7.45 (d, J=7.8 Hz,1H), 7.32 (d, J=7.8 Hz, 1H), 7.17 (t, J=7.5 Hz, 1H), 7.13-7.08 (m, 1H),6.57 (d, J=8.6 Hz, 1H), 5.86 (s, 1H), 5.06 (s, 1H), 4.82-4.77 (m, 1H),3.29 (dd, J=4.8, 15.5 Hz, 1H), 2.99-2.82 (m, 3H), 2.58 (s, 3H),2.32-2.23 (m, 2H), 2.21 (s, 3H); ES-LCMS m/z 391.2 [M+H]⁺.

Step 8:(3R)—N-(8-Isopropyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-122)

To a mixture of(3R)—N-(8-isopropenyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(100 mg, 220.23 mol, 1 eq) in EtOAc (12 mL) was added PtO₂ (250 mg, 1.10mmol, 5.00 eq) under N₂ atmosphere. The mixture was stirred at 15° C.for 20 min under H₂ (15 psi) atmosphere. The mixture was filtered. Thefiltrate was concentrated. The residue was purified by preparative HPLC(column: Agela ASB 150*25 mm*5 m; mobile phase: [water (0.05% HCl)-ACN];B %: 70%-100%, 8 min), followed by lyophilization to yield(3R)—N-(8-isopropyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(50 mg, 116.56 μmol, 52.9% yield, 100% purity, HCl) as a white solid. ¹HNMR (400 MHz, CD₃OD) S ppm 7.90 (s, 1H), 7.37 (d, J=7.6 Hz, 1H), 7.26(d, J=7.6 Hz, 1H), 7.08-7.00 (m, 1H), 6.96 (t, J=7.3 Hz, 1H), 4.73-4.68(m, 1H), 3.23 (dd, J=4.9, 14.7 Hz, 1H), 3.14-3.11 (m, 1H), 2.95 (s, 2H),2.91-2.82 (m, 1H), 2.58 (s, 3H), 2.30 (s, 1H), 2.21 (d, J=6.4 Hz, 1H),1.33 (d, J=6.8 Hz, 6H); ES-LCMS m/z 393.2 [M+H]⁺.

Example 112

Synthesis of I-123

Synthetic Scheme:

Step 1:(3R)—N-[8-(Cyclohexen-1-yl)-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine

A mixture of(3R)—N-[8-bromo-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(200 mg, 408.10 μmol, 1 eq),2-(cyclohexen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (84.94 mg,408.10 μmol, 87.75 μL, 1 eq), Pd(dppf)Cl₂ (29.87 mg, 40.81 μmol, 0.1eq), Cs₂CO₃ (332.47 mg, 1.02 mmol, 2.5 eq) in H₂O (2 mL) and 1,4-dioxane(6 mL) was degassed and purged with N₂ for 3 times, the mixture wasstirred at 90° C. for 2 h under N₂ atmosphere. The reaction mixture wasfiltered and concentrated under reduced pressure to yield a residuewhich was purified by flash silica gel chromatography (from PE/EA=1/0 to1/1, TLC: PE/EtOAc=1/1, R_(f)=0.7) to yield(3R)—N-[8-(cyclohexen-1-yl)-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(110 mg, 193.83 μmol, 47.5% yield, 84.5% purity) as a yellow solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 10.79 (s, 1H), 9.38 (s, 1H), 9.08 (d, J=8.6Hz, 1H), 8.79-8.66 (m, 1H), 8.48-8.37 (m, 1H), 8.28 (s, 1H), 7.35 (d,J=7.8 Hz, 1H), 7.27 (d, J=8.1 Hz, 1H), 7.05-6.97 (m, 1H), 6.96-6.88 (m,1H), 6.73 (br s, 1H), 4.84-4.82 (m, 1H), 3.64 (s, 2H), 3.12 (dd, J=5.1,14.9 Hz, 1H), 3.07-2.83 (m, 3H), 2.24-2.15 (m, 4H), 1.77-1.75 (m, 2H),1.67-1.65 (m, 2H); ES-LCMS m/z 480.3 [M+H]⁺.

Step 2:(3R)—N-[8-Cyclohexyl-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-123)

To a solution of(3R)—N-[8-(cyclohexen-1-yl)-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine (90 mg, 158.59μmol, 1 eq) in EtOAc (5 mL) was added Pd/C (50 mg, 10% purity) under N₂.The suspension was degassed and purged with H₂ several times. Themixture was stirred under H₂ (15 psi) at 10° C. for 12 h. The reactionmixture was filtered and the filtrate was concentrated to yield aresidue which was purified by preparative HPLC (HCl condition; column:Agela ASB 150*25 mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %:85%-100%, 8 min), followed by lyophilization to yield(3R)—N-[8-cyclohexyl-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(16.46 mg, 28.78 μmol, 18.2% yield, 96.9% purity, 2HCl, OR: [α]^(24.6)_(D)=9.235 (7.5 mg/10 mL in MeOH)) as a yellow solid (Optical Rotation:[α]²⁵ _(D)=9.235 (7.5 mg/10 mL in MeOH)). ¹H NMR (400 MHz, DMSO-d₆) δppm 10.79 (s, 1H), 9.37 (s, 1H), 9.02 (d, J=8.6 Hz, 1H), 8.70 (d, J=2.7Hz, 1H), 8.44 (dd, J=1.6, 9.9 Hz, 1H), 8.13 (s, 1H), 7.34 (d, J=7.6 Hz,1H), 7.27 (d, J=7.8 Hz, 1H), 7.07-6.97 (m, 1H), 6.96-6.88 (m, 1H),4.90-4.75 (m, 1H), 3.15-3.07 (m, 1H), 3.05-2.80 (m, 4H), 2.20-2.19 (m,2H), 1.82-1.79 (m, 2H), 1.87-1.69 (m, 3H), 1.62-1.59 (m, 2H), 1.44-1.41(m, 2H), 1.31-1.22 (m, 1H); ES-LCMS m/z 482.3 [M+H]⁺.

Example 113

Synthesis of I-124

Synthetic Scheme:

Step 1: 3-Methylpyrazolo[1,5-a]pyrimidine-5,7-diol

To a solution of 4-methyl-1H-pyrazol-5-amine (2 g, 20.59 mmol, 1 eq) anddimethyl propanedioate (2.86 g, 21.62 mmol, 2.48 mL, 1.05 eq) in MeOH(20 mL) was added a solution of Na (946.88 mg, 41.19 mmol, 976.16 μL,2.0 eq) in MeOH (20 mL) under N₂ at 20° C. The mixture was stirred at80° C. for 12 h. The reaction mixture was cooled to 20° C., filtered andcollected the white solid. The solid was diluted with 1N HCl (20 mL) andstirred for 10 min, filtered and the solid was dried under reducedpressure to yield product of 3-methylpyrazolo[1,5-a]pyrimidine-5,7-diol(2 g, 12.11 mmol, 58.8% yield, crude purity) as a white solid which wasused in the next step without further purification. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 10.54 (br s, 1H), 7.31 (s, 1H), 4.21 (br s, 1H), 1.96 (s,3H).

Step 2: 5,7-Dichloro-3-methyl-pyrazolo[1,5-a]pyrimidine

A solution of 3-methylpyrazolo[1,5-a]pyrimidine-5,7-diol (500 mg, 3.03mmol, 1 eq) in POCl₃ (5 mL) was stirred at 110° C. for 3 h. The reactionmixture was concentrated under reduced pressure. The residue was dilutedDCM (20 mL×2) and concentrated under reduced pressure. The residue wasdiluted DCM (20 mL), adjusted to pH to 9-10 with DIEA and concentratedunder reduced pressure to yield a residue which was purified on silicagel column chromatography (from PE/EtOAc=1/0 to 5/1, TLC: PE/EtOAc=10/1,R_(f)=0.68) to yield product of5,7-dichloro-3-methyl-pyrazolo[1,5-a]pyrimidine (400 mg, 1.98 mmol,65.3% yield, 99.8% purity) as a white solid. ¹H NMR (400 MHz, CDCl₃) δppm 8.09 (s, 1H), 6.93 (s, 1H), 2.37 (s, 3H); ES-LCMS m/z 202.0, 204.0[M+H]⁺.

Step 3:5-Chloro-7-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine

5,7-Dichloro-3-methyl-pyrazolo[1,5-a]pyrimidine (190 mg, 938.52 μmol, 1eq), (5-fluoro-3-pyridyl)boronic acid (138.86 mg, 985.45 μmol, 1.05 eq),Pd(dppf)Cl₂ (68.67 mg, 93.85 μmol, 0.1 eq) and Cs₂CO₃ (611.58 mg, 1.88mmol, 2 eq) were taken up into a microwave tube in 1,4-dioxane (3 mL)and H₂O (1 mL). The sealed tube was heated at 110° C. for 1 h undermicrowave. The mixture was concentrated and water (10 mL) was added. Themixture was extracted with EtOAc (20 mL×3). The combined organic layerswere dried over Na₂SO₄, filtered and concentrated to yield a residuewhich was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1, R_(f)=0.50) to yield5-chloro-7-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (130mg, 494.92 μmol, 52.7% yield, 100.0% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.97 (s, 1H), 8.69 (d, J=2.7 Hz, 1H), 8.38-8.31(m, 1H), 8.04 (s, 1H), 6.92 (s, 1H), 2.40 (s, 3H); ES-LCMS m/z 263.0,265.0 [M+H]⁺.

Step 4:7-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-5-amine (I-124)

5-Chloro-7-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (70mg, 266.49 μmol, 1 eq), 2-(1H-indol-3-yl)ethanamine (64.04 mg, 399.74μmol, 1.5 eq) and DIEA (103.33 mg, 799.48 μmol, 139.26 μL, 3 eq) weretaken up into a microwave tube in i-PrOH (2 mL). The sealed tube washeated at 150° C. for 3 h under microwave. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Agela Durashell C18 150*25 5 u;mobile phase: [water (0.05% HCl)-ACN]; B %: 32%-55%, 9 min), followed bylyophilization to yield7-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-5-amine(28.21 mg, 59.77 μmol, 22.4% yield, 97.3% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD+Na₂CO₃) δ ppm 8.80 (br s, 1H), 8.62 (d,J=2.7 Hz, 1H), 8.18 (br s, 1H), 7.74-7.65 (m, 2H), 7.31 (d, J=8.1 Hz,1H), 7.12-7.04 (m, 2H), 7.03-6.97 (m, 1H), 6.27 (br s, 1H), 3.78 (t,J=7.2 Hz, 2H), 3.11 (t, J=7.3 Hz, 2H), 2.24 (s, 3H); ES-LCMS m/z 387.1[M+H]⁺.

Example 114

Synthesis of I-125

Synthetic Scheme:

Step 1: 5-Fluoro-N′-(1H-imidazol-2-yl)pyridine-3-carboxamidine

To a mixture of ethyl 5-fluoropyridine-3-carboximidate (1 g, 5.65 mmol,1 eq) and 1H-imidazol-2-amine (563.27 mg, 6.78 mmol, 1.2 eq) in ACN (15mL) was added AcONa (926.84 mg, 11.30 mmol, 2 eq). The mixture wasstirred at 80° C. for 25 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified on silica gel column chromatography (from PE/EtOAc=1/0 to 2/1,TLC: PE/EtOAc=2/1, R_(f)=0.35) to yield5-fluoro-N-(1H-imidazol-2-yl)pyridine-3-carboxamidine (400 mg, 1.75mmol, 31.1% yield, 90.0% purity) as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 11.67 (br s, 1H), 9.97 (br s, 1H), 9.05 (s, 1H), 8.70 (d,J=2.4 Hz, 1H), 8.29-8.12 (m, 2H), 6.92 (s, 1H), 6.85 (s, 1H); ES-LCMSm/z 206.1 [M+H]⁺.

Step 2: 2-(5-Fluoro-3-pyridyl)imidazo[1,2-a][1,3,5]triazin-4-ol

To a solution of 5-fluoro-N′-(1H-imidazol-2-yl)pyridine-3-carboxamidine(400 mg, 1.75 mmol, 1 eq) in THE (5 mL) and 1,4-dioxane (5 mL) was addeddiphosgene (694.18 mg, 3.51 mmol, 423.28 μL, 2 eq). The mixture wasstirred at 80° C. for 2 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to yield a residue which was washedwith DMF (5 mL), filtered and concentrated to yield2-(5-fluoro-3-pyridyl)imidazo[1,2-a][1,3,5]triazin-4-ol (250 mg, 973.24μmol, 55.5% yield, 90.0% purity) as a white solid which was used in thenext step without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm13.64 (br s, 1H), 9.29 (s, 1H), 8.78 (d, J=2.7 Hz, 1H), 8.35 (J=10.0 Hz,1H), 7.73-7.65 (m, 2H); ES-LCMS m/z 232.0 [M+H]⁺.

Step 3: 4-Chloro-2-(5-fluoro-3-pyridyl)imidazo[1,2-a][1,3,5]triazine

To a solution of 2-(5-fluoro-3-pyridyl)imidazo[1,2-a][1,3,5]triazin-4-ol(100 mg, 389.30 μmol, 1 eq) in POCl₃ (12 mL) was added DIEA (1.48 g,11.48 mmol, 2 mL, 29.49 eq). The mixture was stirred at 120° C. for 3 hunder N₂ atmosphere. The reaction mixture was concentrated under reducedpressure to yield4-chloro-2-(5-fluoro-3-pyridyl)imidazo[1,2-a][1,3,5]triazine (90 mg,360.53 μmol, 92.6% yield, crude) as a black solid which was used in thenext step without further purification. ES-LCMS m/z 350.1 [M+H]⁺.

Step 4:(3R)—N-[2-(5-Fluoro-3-pyridyl)imidazo[1,2-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-125)

To a mixture of4-chloro-2-(5-fluoro-3-pyridyl)imidazo[1,2-a][1,3,5]triazine (90 mg,360.53 μmol, 1 eq) and (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(80.58 mg, 432.64 μmol, 1.2 eq) in ACN (10 mL) was added DIEA (5.34 g,41.34 mmol, 7.20 mL, 114.65 eq). The mixture was stirred at 80° C. for 2h under N₂ atmosphere. The reaction mixture was concentrated to yield aresidue which was purified by preparative HPLC (column: Agela DuraShell150 mm_25 mm_5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 23%-53%,8.5 min), followed by lyophilization to yield a product which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 μm;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 40%-70%, 8min), followed by lyophilization to yield(3R)—N-[2-(5-fluoro-3-pyridyl)imidazo[1,2-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(15.11 mg, 37.83 μmol, 10.5% yield, 100.0% purity, [α]²⁴ _(D)=98.97(MeOH, c=0.050 g/100 mL)) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δppm 9.39 (s, 1H), 8.55-8.47 (m, 2H), 7.93 (d, J=1.7 Hz, 1H), 7.56 (s,1H), 7.36 (d, J=7.6 Hz, 1H), 7.25 (d, J=8.1 Hz, 1H), 7.05-6.98 (m, 1H),6.97-6.90 (m, 1H), 4.85-4.79 (m, 1H), 3.36 (J=5.1 Hz, 1H), 3.08-2.83 (m,3H), 2.44 (J=9.8 Hz, 1H), 2.26-2.14 (m, 1H); ES-LCMS m/z 400.2 [M+H]⁺.

Example 115

Synthesis of I-126

Synthetic Scheme:

Step 1:(3R)—N-(2-Chloropyrido[3,2-d]pyrimidin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine

To a solution of 2,4-dichloropyrido[3,2-d]pyrimidine (100 mg, 499.94μmol, 1 eq) in THE (3 mL) was added DIEA (193.84 mg, 1.50 mmol, 261.24μL, 3 eq) and (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (93.11 mg,499.94 μmol, 1 eq). The mixture was stirred at 60° C. for 3 h. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by flash silica gel chromatography (fromPE/EtOAC=100/1 to 1/1, TLC: PE/EtOAc=1/1, R_(f)=0.5) to yield(3R)—N-(2-chloropyrido[3,2-d]pyrimidin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(150 mg, 428.80 μmol, 85.8% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 8.72 (dd, J=1.5, 4.4 Hz, 1H), 7.95 (dd,J=1.5, 8.3 Hz, 1H), 7.75 (dd, J=4.2, 8.6 Hz, 1H), 7.37 (d, J=7.6 Hz,1H), 7.26 (d, J=8.1 Hz, 1H), 7.07-7.00 (m, 1H), 6.98-6.92 (m, 1H),4.80-4.68 (m, 1H), 3.21 (dd, J=5.1, 15.2 Hz, 1H), 3.06-2.80 (m, 3H),2.37-2.12 (m, 2H); ES-LCMS m/z 350.1 [M+H]⁺.

Step 2:(3R)—N-[2-(5-Fluoro-3-pyridyl)pyrido[3,2-d]pyrimidin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-126)

(3R)—N-(2-chloropyrido[3,2-d]pyrimidin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(60 mg, 171.52 μmol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (29.00 mg,205.82 μmol, 1.2 eq), Pd(dppf)Cl₂ (12.55 mg, 17.15 μmol, 0.1 eq) andCs₂CO₃ (167.65 mg, 514.56 μmol, 3 eq) were taken up into a microwavetube in 1,4-dioxane (2 mL) and H₂O (0.5 mL). The sealed tube was heatedat 110° C. for 0.5 h under microwave. The reaction mixture was dilutedwith water (5 mL) and extracted with EtOAc (10 mL×3). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by preparativeHPLC (column: Agela ASB 150*25 mm*5 um; mobile phase: [water(0.05%HCl)-ACN]; B %: 55%-85%, 8 min), followed by lyophilization to yield(3R)—N-[2-(5-fluoro-3-pyridyl)pyrido[3,2-d]pyrimidin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(39.09 mg, 95.24 μmol, 55.5% yield, 100.0% purity, OR: [α]^(23.6)_(D)=0.471 (MeOH, c=0.110 g/100 mL).) as a yellow solid. ¹H NMR (400MHz, CD₃OD) δ ppm 9.32 (s, 1H), 9.01 (dd, J=1.2, 4.4 Hz, 1H), 8.83 (d,J=2.4 Hz, 1H), 8.61-8.49 (m, 1H), 8.32 (dd, J=1.2, 8.6 Hz, 1H), 8.04(dd, J=4.4, 8.6 Hz, 1H), 7.33 (d, J=7.6 Hz, 1H), 7.25 (d, J=8.1 Hz, 1H),6.98 (t, J=7.5 Hz, 1H), 6.91-6.81 (m, 1H), 5.22-5.04 (m, 1H), 3.28 (s,1H), 3.12-2.86 (m, 3H), 2.43-2.27 (m, 2H); ES-LCMS m/z 411.2 [M+H]⁺.

Example 116

Synthesis of I-128a

Synthetic Scheme:

Step 1:(6S)—N6-[2-(5-Fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-4,5,6,7-tetrahydro-1,3-benzothiazole-2,6-diamine(I-128a)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(70 mg, 236.12 μmol, 1 eq),(6S)-4,5,6,7-tetrahydro-1,3-benzothiazole-2,6-diamine (43.96 mg, 259.74μmol, 1.1 eq) and DIEA (91.55 mg, 708.37 μmol, 123.39 μL, 3 eq) ini-PrOH (10 mL) was stirred at 80° C. for 2 h. The mixture wasconcentrated under reduced pressure to give a residue which was washedwith MeOH (20 mL×2) to yield(6S)—N6-[2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-4,5,6,7-tetrahydro-1,3-benzothiazole-2,6-diamine(22.62 mg, 53.29 μmol, 22.6% yield, 100.0% purity, SFC: Rt=5.691 min,ee=99.8%, OR: [α]^(21.9) _(D)=−34.176 (CHCl₃, c=0.104 g/100 mL)) as awhite solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.37 (s, 1H), 8.97 (br s,1H), 8.71 (d, J=2.8 Hz, 1H), 8.44 (d, J=10.0 Hz, 1H), 8.13 (s, 1H), 6.72(s, 2H), 4.75-4.72 (m, 1H), 3.49-3.19 (m, 1H), 2.90-2.83 (m, 2H),2.65-2.58 (m, 1H), 2.54-2.50 (m, 1H), 2.09-2.05 (m, 2H), 1.36 (d, J=6.8Hz, 6H); ES-LCMS m/z 425.2 [M+H]⁺

Example 117

Synthesis of I-128b

Synthetic Scheme:

Step 1:(6R)—N6-[2-(5-Fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-4,5,6,7-tetrahydro-1,3-benzothiazole-2,6-diamine(I-128b)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(70 mg, 236.12 μmol, 1 eq),(6R)-4,5,6,7-tetrahydro-1,3-benzothiazole-2,6-diamine (43.96 mg, 259.74μmol, 1.1 eq) and DIEA (91.55 mg, 708.37 μmol, 123.38 μL, 3 eq) ini-PrOH (10 mL) was stirred at 80° C. for 2 h. The mixture wasconcentrated under reduced pressure to yield a residue which was washedwith MeOH (20 mL×2) to yield(6R)—N6-[2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-4,5,6,7-tetrahydro-1,3-benzothiazole-2,6-diamine(20.56 mg, 48.43 μmol, 20.5% yield, 100.0% purity, SFC: R_(t)=7.228 min,ee=98.44%, OR: [α]^(22.1) _(D)=29.753 (CHCl₃, c=0.100 g/100 mL)) as awhite solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.37 (s, 1H), 8.97 (d,J=8.4 Hz, 1H), 8.71 (d, J=2.8 Hz, 1H), 8.46-8.42 (m, 1H), 8.13 (s, 1H),6.72 (s, 2H), 4.77-4.72 (m, 1H), 3.23-3.17 (m, 1H), 2.90-2.83 (m, 2H),2.67-2.64 (m, 1H), 2.58-2.53 (m, 1H), 2.07-2.05 (m, 2H), 1.36 (d, J=6.8Hz, 6H); ES-LCMS m/z 425.2 [M+H]⁺.

Example 118

Synthesis of I-129

Synthetic Scheme:

Step 1:N′-[4-(Dimethylamino)-1H-pyrazol-5-yl]-5-fluoro-pyridine-3-carboxamidine

To a solution of N4,N4-dimethyl-1H-pyrazole-4,5-diamine (1.2 g, 6.27mmol, 1 eq, HCl) in ACN (20 mL) was added ethyl5-fluoropyridine-3-carboximidate (1.11 g, 6.27 mmol, 1 eq). The mixturewas stirred at 60° C. for 12 h under N₂ atmosphere. The reaction mixturewas concentrated under reduced pressure to yield a residue which waspurified on silica gel column chromatography (from DCM/MeOH=1/0 to 5/1,TLC: DCM/MeOH=10/1, R_(f)=0.3) to yieldN-[4-(dimethylamino)-1H-pyrazol-5-yl]-5-fluoro-pyridine-3-carboxamidine(700 mg, 2.52 mmol, 40.2% yield, 89.5% purity) as a yellow solid. ¹H NMR(400 MHz, DMSO-d₆) δ ppm 11.96 (br s, 1H), 9.07 (s, 1H), 8.66 (d, J=2.0Hz, 1H), 8.17 (d, J=10.0 Hz, 1H), 7.18 (s, 1H), 2.72 (br s, 6H); ES-LCMSm/z 248.9 [M+H]⁺.

Step 2:2-8-(Dimethylamino)-2-(5-Fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-ol

To a solution ofN-[4-(dimethylamino)-1H-pyrazol-5-yl]-5-fluoro-pyridine-3-carboxamidine(160 mg, 576.82 μmol, 1 eq) in THE (1 mL) and toluene (8 mL) was addeddiphosgene (342.34 mg, 1.73 mmol, 208.74 μL, 3 eq) under N₂ at 15° C.,the mixture was stirred at 15° C. for 0.5 h. Then the mixture wasstirred at 130° C. for 2 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to yield8-(dimethylamino)-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-ol(150 mg, crude) as a yellow solid which was used in the next stepwithout further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 9.15 (s,1H), 9.07 (s, 1H), 8.92 (d, J=2.3 Hz, 1H), 8.79 (d, J=2.8 Hz, 1H), 8.03(s, 1H), 3.03 (s, 6H); ES-LCMS m/z 275.1 [M+H]⁺.

Step 34-Chloro-2-(5-fluoro-3-pyridyl)-N,N-dimethyl-pyrazolo[1,5-a][1,3,5]triazin-8-amine

A mixture of8-(dimethylamino)-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-ol(150 mg, 546.94 μmol, 1 eq) in POCl₃ (9 g, 58.70 mmol, 5.45 mL, 107.32eq) was degassed and purged with N₂ for 3 times, stirred at 120° C. for1 h under N₂ atmosphere. The reaction mixture was concentrated underreduced pressure to yield a residue which was quenched by addition ofwater (50 mL), adjusted pH to 8 by aq. NaHCO₃, extracted with EtOAc (30mL×3). The combined organic layers were washed with brine (10 mL), driedover Na₂SO₄, filtered and concentrated under reduced pressure to yield aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1, R_(f)=0.31) to yield4-chloro-2-(5-fluoro-3-pyridyl)-N,N-dimethyl-pyrazolo[1,5-a][1,3,5]triazin-8-amine(25 mg, 76.87 μmol, 14.0% yield, 90% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.35 (s, 1H), 8.51 (br s, 1H), 8.27 (d, J=8.8 Hz,1H), 7.85 (s, 1H), 3.05 (s, 6H); ES-LCMS m/z 293.1, 295.0 [M+H]⁺.

Step 4:2-(5-Fluoro-3-pyridyl)-N8,N8-dimethyl-N4-[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]pyrazolo[1,5-a][1,3,5]triazine-4,8-diamine(I-129)

To a solution of4-chloro-2-(5-fluoro-3-pyridyl)-N,N-dimethyl-pyrazolo[1,5-a][1,3,5]triazin-8-amine (25 mg, 76.87 μmol, 1 eq) in i-PrOH (3 mL) was added(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (17.18 mg, 92.24 μmol, 1.2eq). The mixture was stirred at 40° C. for 2 h. The reaction mixture wasconcentrated to yield a residue which was purified by preparative HPLC(column: Agela ASB 150*25 mm*5 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 38%-68%, 8 min) twice, followed by lyophilization toyield2-(5-fluoro-3-pyridyl)-N8,N8-dimethyl-N4-[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]pyrazolo[1,5-a][1,3,5]triazine-4,8-diamine(34.45 mg, 62.30 μmol, 81.0% yield, 99.8% purity, 3HCl, [α]^(20.6)_(D)=+36.080 (MeOH, c=0.1 g/100 mL)) as a yellow solid. ¹H NMR (400 MHz,CD₃OD) δ ppm 9.51 (br s, 1H), 8.74 (m, 2H), 8.42 (br s, 1H), 7.37 (d,J=7.8 Hz, 1H), 7.28 (d, J=8.3 Hz, 1H), 7.04 (t, J=7.5 Hz, 1H), 6.99-6.92(m, 1H), 5.00-4.92 (m, 1H), 3.53 (br s, 6H), 3.28-2.89 (m, 4H),2.44-2.23 (m, 2H); ES-LCMS m/z 443.2 [M+H]⁺.

Example 119

Synthesis of I-130

Synthetic Scheme:

Step 1: tert-Butyl(3R)-3-[[8-bromo-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

A mixture of(3R)—N-[8-bromo-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(560 mg, 1.09 mmol, 1 eq), Boc₂O (1.43 g, 6.57 mmol, 1.51 mL, 6 eq) andDMAP (534.93 mg, 4.38 mmol, 4 eq) in 1,4-dioxane (50 mL) was stirred at110° C. for 12 h. TLC (PE/EtOAc=5/1, R_(f)=0.65) showed the startingmaterial was consumed completely. The reaction mixture was concentratedunder reduced pressure to yield a residue which was purified by flashsilica gel chromatography (from PE/EtOAc=1/0 to 10/1, TLC: PE/EtOAc=5/1,R_(f)=0.65) to yield tert-butyl(3R)-3-[[8-bromo-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(660 mg, 972.66 μmol, 88.9% yield, 100.0% purity) as a white solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.49 (s, 1H), 8.60 (d, J=2.8 Hz, 1H), 8.44(d, J=8.4 Hz, 1H), 8.23 (s, 1H), 8.10 (d, J=8.4 Hz, 1H), 7.33 (d, J=7.8Hz, 1H), 7.22-7.16 (m, 2H), 4.92 (br s, 1H), 3.34 (d, J=16.8 Hz, 1H),3.25-3.11 (m, 3H), 2.50 (m, 1H), 2.33 (m, 1H), 1.67 (s, 9H), 1.39 (s,9H); ES-LCMS m/z 678.2, 680.2 [M+H]⁺.

Step 2:2-(5-Fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carbonitrile(I-130)

A mixture of tert-butyl(3R)-3-[[8-bromo-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(50 mg, 72.36 μmol, 1 eq), Zn(CN)₂ (33.99 mg, 289.44 μmol, 18.37 μL, 4eq) and Pd(PPh₃)₄ (16.72 mg, 14.47 μmol, 0.2 eq) in DMF (3 mL) weretaken up into a microwave tube and then purged with N₂ for 1 min. Thesealed tube was stirred at 170° C. for 20 min under microwave (1 bar).The reaction mixture was extracted with EtOAc (20 mL×3). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated toyield a residue which was purified by preparative HPLC (column: AgelaASB 150*25 mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 55%-85%,8 min), followed by lyophilization to yield2-(5-fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carbonitrile(15.36 mg, 29.98 μmol, 41.4% yield, 97.1% purity, 2HCl, [α]¹⁸¹_(D)=45.287, (MeOH, c=0.078 g/100 mL)) as a yellow solid. ¹H NMR (400MHz, CD₃OD) δ ppm 9.48 (s, 1H), 8.80-8.71 (m, 2H), 8.43 (s, 1H), 7.32(d, J=7.6 Hz, 1H), 7.24 (d, J=8.1 Hz, 1H), 7.00 (t, J=7.5 Hz, 1H),6.94-6.88 (m, 1H), 4.83 (m, 1H), 3.26 (m, 1H), 3.08-2.88 (m, 3H),2.37-2.35 (m, 1H), 2.31-2.18 (m, 1H); ES-LCMS m/z 425.2 [M+H]⁺.

Example 120

Synthesis of I-131

Synthetic Scheme:

Step 1:N-(2-(1H-Indol-3-yl)ethyl)-8-(tert-butyl)-2-(5-fluoropyridin-3-yl)pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-131)

To a solution of8-tert-butyl-4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine(60 mg, 176.62 μmol, 1 eq) in i-PrOH (3 mL) was added2-(1H-indol-3-yl)ethanamine (29.43 mg, 183.68 μmol, 1.04 eq) and DIPEA(114.13 mg, 883.10 μmol, 153.82 □μL, 5 eq). The reaction mixture wasstirred at 50° C. for 3 h. The reaction mixture was concentrated underreduced pressure to give a residue which was purified by preparativeHPLC (column: Agela Durashell C18 150*25 5 u; mobile phase: [water(0.05% HCl)-ACN]; B %: 75%-95%, 8 min), followed by lyophilization toyield8-tert-butyl-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(27.49 mg, 54.18 μmol, 30.7% yield, 99.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.13 (s, 1H), 8.80 (s, 1H), 8.55(d, J=8.6 Hz, 1H), 7.95 (s, 1H), 7.69-7.62 (m, 1H), 7.19-7.12 (m, 1H),7.01-6.93 (m, 3H), 4.02 (J=6.7 Hz, 2H), 3.16 (t, J=6.7 Hz, 2H), 1.48 (s,9H); ES-LCMS m/z 430.3 [M+H]⁺.

Example 121

Synthesis of I-132

Step 1: (Z)—N-(4-(tert-Butyl)-1H-pyrazol-5-yl)-5-fluoronicotinimidamide

To a solution of ethyl 5-fluoropyridine-3-carboximidate (400 mg, 2.26mmol, 1 eq) in toluene (15 mL) was added 4-tert-butyl-1H-pyrazol-5-amine(314.54 mg, 2.26 mmol, 1 eq). The mixture was stirred at 130° C. for 24h under N₂. The reaction mixture was concentrated under reduced pressureto yield a residue which was purified on silica gel columnchromatography (from PE/EtOAc=1/0 to 5/1, TLC: PE/EtOAc=1/1, R_(f)=0.23)to yieldN′-(4-tert-butyl-1H-pyrazol-5-yl)-5-fluoro-pyridine-3-carboxamidine (400mg, 1.45 mmol, 64.4% yield, 95.0% purity) as a white solid. ¹H NMR (400MHz, CDCl₃) δ ppm 9.59 (s, 1H), 8.94 (s, 1H), 8.57-8.48 (m, 1H), 8.02(d, J=9.5 Hz, 1H), 7.23 (s, 1H), 1.41 (s, 9H); ES-LCMS m/z 262.1 [M+H]⁺.

Step 2:8-(tert-Butyl)-2-(5-fluoropyridin-3-yl)pyrazolo[1,5-a][1,3,5]triazin-4-ol

To a solution ofN-(4-tert-butyl-1H-pyrazol-5-yl)-5-fluoro-pyridine-3-carboxamidine (400mg, 1.45 mmol, 1 eq) in 1,4-dioxane (5 mL) and THF (5 mL) was addeddiphosgene (575.40 mg, 2.91 mmol, 350.85 μL, 2 eq). The mixture wasstirred at 80° C. for 7 h under N₂. The reaction mixture wasconcentrated under reduced pressure to yield8-tert-butyl-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-ol(400 mg, 1.32 mmol, 91.0% yield, 95.0% purity) as a yellow solid whichwas used in next step without further purification. ¹H NMR (400 MHz,CD₃OD) δ ppm 9.22 (s, 1H), 8.89 (s, 1H), 8.55 (d, J=9.0 Hz, 1H), 8.03(s, 1H), 1.49 (s, 9H); ES-LCMS m/z 287.8 [M+H]⁺.

Step 3:8-(tert-Butyl)-4-chloro-2-(5-fluoropyridin-3-yl)pyrazolo[1,5-a][1,3,5]triazine

A solution of8-tert-butyl-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-ol(400 mg, 1.32 mmol, 1 eq) in POCl₃ (2 g, 13.04 mmol, 1.21 mL, 9.86 eq)was stirred at 110° C. for 2 h under N₂ atmosphere. The reaction mixturewas concentrated under reduced pressure. The residue was dissolved inice-water (10 mL) and extracted with DCM (10 mL×3). The organic layerwas dried over Na₂SO₄, filtered and concentrated to yield a residuewhich was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 5/1, TLC: PE/EtOAc=3/1, R_(f)=0.7) to yield8-tert-butyl-4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine(130 mg, 382.68 μmol, 28.9% yield, 90.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.44 (s, 1H), 8.56 (s, 1H), 8.35 (d, J=9.0Hz, 1H), 8.11 (s, 1H), 1.47 (s, 9H); ES-LCMS m/z 305.7 [M+H]⁺.

Step 4:(R)—N-(8-(tert-Butyl)-2-(5-fluoropyridin-3-yl)pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-132)

To a solution of8-tert-butyl-4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine(60 mg, 176.62 μmol, 1 eq) in i-PrOH (3 mL) was added(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (34.21 mg, 183.69 μmol, 1.04eq) and DIPEA (114.14 mg, 883.11 μmol, 153.82 μL, 5 eq). The reactionmixture was stirred at 50° C. for 3 h. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Agela Durashell C18 150*25 5 u;mobile phase: [water (0.05% HCl)-ACN]; B %: 75%-95%, 8 min), followed bylyophilization to yield(3R)—N-[8-tert-butyl-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(28.87 mg, 53.29 μmol, 30.2% yield, 97.6% purity, 2HCl, [α]²¹_(D)=18.883 (MeOH, c=0.100 g/100 mL)) as a yellow solid. ¹H NMR (400MHz, CD₃OD) δ ppm 9.41 (s, 1H), 8.69-8.61 (m, 2H), 7.94 (s, 1H), 7.35(d, J=7.8 Hz, 1H), 7.25 (d, J=8.1 Hz, 1H), 7.01 (t, J=7.5 Hz, 1H),6.97-6.87 (m, 1H), 4.81 (m, 1H), 3.25 (d, J=4.9 Hz, 1H), 3.10-2.84 (m,3H), 2.35 (m, 1H), 2.29-2.18 (m, 1H), 1.50 (s, 9H); ES-LCMS m/z 456.3[M+H]⁺.

Example 122

Synthesis of I-133

Synthetic Scheme:

Step 1: 3-(2-Aminoethyl)-1H-indol-7-ol

To a solution of 2-(7-benzyloxy-1H-indol-3-yl)ethanamine (150 mg, 563.19μmol, 1 eq) in MeOH (10 mL) was added Pd/C (50 mg, 10%). The mixture wasdegassed and purged with H₂ for 3 times and stirred at 50° C. for 1 h.The mixture was filtered and concentrated to yield3-(2-aminoethyl)-1H-indol-7-ol (90 mg, 510.74 μmol, 90.7% yield) asyellow solid which was used in the next step without furtherpurification. ¹H NMR (400 MHz, CD₃OD) δ ppm 7.06-7.00 (m, 2H), 6.85-6.77(m, 1H), 6.50 (d, J=7.6 Hz, 1H), 2.96-2.83 (m, 4H).

Step 2:3-[2-[[5-(5-Fluoro-3-pyridyl)-3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-1H-indol-7-ol (I-133)

A mixture of[7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-3-yl]methanol(55 mg, 171.12 μmol, 1 eq), 3-(2-aminoethyl)-1H-indol-7-ol (39.20 mg,222.45 μmol, 1.3 eq) and DIEA (66.35 mg, 513.35 μmol, 89.42 μL, 3 eq) ini-PrOH (10 mL) was degassed and purged with N₂ for 3 times. The mixturewas stirred at 80° C. for 5 h under N₂ atmosphere. The reaction mixturewas concentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Xtimate C18 150*25 mm*5 μm; mobilephase: [Water-ACN]; B %: 17%-47%, 8.5 min), followed by lyophilizationto yield3-[2-[[5-(5-fluoro-3-pyridyl)-3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-1H-indol-7-ol(16.9 mg, 38.27 μmol, 22.4% yield, 94.8% purity) as a gray solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 8.67 (s, 1H), 8.47 (d, J=2.4 Hz, 1H), 8.06 (s,1H), 7.73-7.67 (m, 1H), 7.21 (d, J=8.1 Hz, 1H), 6.94-6.87 (m, 2H), 6.51(d, J=7.3 Hz, 1H), 5.96 (s, 1H), 4.80 (s, 2H), 3.88 (t, J=6.1 Hz, 2H),3.21-3.11 (m, 2H); ES-LCMS m/z 419.1 [M+H]⁺.

Example 123

Synthesis of I-135

Synthetic Scheme:

Step 1:(3R)—N-(4-Chloro-6-isopropoxy-1,3,5-triazin-2-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine

To a solution of 2,4-dichloro-6-isopropoxy-1,3,5-triazine (150 mg,648.90 μmol, 1 eq) in THE (5 mL) was added DIPEA (119.93 mg, 927.92μmol, 161.63 □μL, 1.43 eq) and(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (125.69 mg, 674.85 μmol,1.04 eq) in THF (5 mL) dropwise at 0° C. The mixture was stirred at 0°C. for 1 h and concentrated under reduced pressure to yield a residuewhich was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=1/1, R_(f)=0.47) to yield(3R)—N-(4-chloro-6-isopropoxy-1,3,5-triazin-2-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(220 mg, 553.33 μmol, 85.3% yield, 90.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 7.80 (d, J=6.4 Hz, 1H), 7.45-7.38 (m, 1H),7.29 (dd, J=3.1, 7.7 Hz, 1H), 7.18-7.05 (m, 2H), 6.06-5.59 (m, 1H),5.41-4.98 (m, 1H), 4.65-4.51 (m, 1H), 3.16 (d, J=15.4 Hz, 1H), 2.96-2.66(m, 3H), 2.18-2.06 (m, 2H), 1.31 (d, J=5.1 Hz, 6H); ES-LCMS m/z 358.1[M+H]⁺.

Step 2:(3R)—N-[4-(5-Fluoro-3-pyridyl)-6-isopropoxy-1,3,5-triazin-2-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-135)

To a solution of(3R)—N-(4-chloro-6-isopropoxy-1,3,5-triazin-2-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(200 mg, 503.02 μmol, 1 eq) in 1,4-dioxane (3 mL) and H₂O (1 mL) wasadded (5-fluoro-3-pyridyl)boronic acid (70.88 mg, 503.02 μmol, 1 eq),Pd(dppf)Cl₂ (36.81 mg, 50.30 μmol, 0.1 eq) and Cs₂CO₃ (491.68 mg, 1.51mmol, 3 eq) under N₂. The mixture was stirred at 100° C. for 3 h underN₂. The reaction mixture was concentrated to yield a residue which waspurified by preparative HPLC (HCl condition; column: Agela ASB 150*25mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 60%-90%, 8 min) toyield(3R)—N-[4-(5-fluoro-3-pyridyl)-6-isopropoxy-1,3,5-triazin-2-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(55.56 mg, 113.07 μmol, 22.5% yield, 100.0% purity, 2HCl, [α]^(21.2)_(D)=56.652 (MeOH, c=0.100 g/100 mL)) as a yellow solid. ¹H NMR (400MHz, CD₃OD) δ ppm 9.26 (s, 1H), 8.75 (s, 1H), 8.50 (d, J=7.6 Hz, 1H),7.30 (d, J=7.6 Hz, 1H), 7.20 (d, J=7.8 Hz, 1H), 7.03-6.86 (m, 2H), 5.44(s, 1H), 4.65-4.37 (m, 1H), 3.16 (d, J=14.7 Hz, 1H), 2.89 (s, 2H), 2.70(dd, J=8.8, 14.2 Hz, 1H), 2.24 (s, 1H), 2.03 (s, 1H), 1.41 (d, J=5.1 Hz,6H); ES-LCMS m/z 419.0 [M+H]⁺.

Example 124

Synthesis of I-136

Synthetic Scheme:

Step 1: 2,4-Dichloro-6-isopropoxy-1,3,5-triazine

To a solution of i-PrOH (322.62 mg, 5.37 mmol, 410.98 μL, 1.1 eq) in THE(12 mL) was added NaH (253.76 mg, 6.34 mmol, 60% purity, 1.3 eq) at 0°C. under N₂. The mixture was stirred at 0° C. for 0.5 h. Then2,4,6-trichloro-1,3,5-triazine (1 g, 4.88 mmol, 1 eq) in THE (8 mL) wasadded to the mixture dropwise at 0° C. The mixture was stirred at 10° C.for 11.5 h under N₂. TLC (PE/EtOAc=6:1, R_(f)=0.6) showed the reactionwas completed. The reaction mixture was dissolved in water (50 mL) at 0°C. slowly. The mixture was extracted with EtOAc (50 mL×3). The combinedorganic layers were washed with brine (50 mL), dried over Na₂SO₄,filtered and concentrated to yield a residue which was purified onsilica gel column chromatography (from PE/EtOAc=1/0 to 20/1, TLC:PE/EtOAc=6/1, R_(f)=0.6) to yield2,4-dichloro-6-isopropoxy-1,3,5-triazine (430 mg, 1.86 mmol, 38.1%yield, 90.0% purity) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ ppm5.40-5.25 (m, 1H), 1.37 (d, J=6.1 Hz, 6H).

Step 2:4-Chloro-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-1,3,5-triazin-2-amine

To a solution of 2,4-dichloro-6-isopropoxy-1,3,5-triazine (100 mg,432.60 μmol, 1 eq) in THF (10 mL) was added DIPEA (79.95 mg, 618.62μmol, 107.75 μL, 1.43 eq) and 2-(1H-indol-3-yl)ethanamine (72.08 mg,449.90 umol, 1.04 eq) in THE (5 mL) dropwise at 0° C. The mixture wasstirred at 0° C. for 1 h and concentrated under reduced pressure toyield a residue which was purified on silica gel column chromatography(from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=1/1, R_(f)=0.47) to yield4-chloro-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-1,3,5-triazin-2-amine(90 mg, 244.12 μmol, 56.4% yield, 90.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.13-7.96 (m, 1H), 7.59-7.47 (m, 1H),7.33-7.24 (m, 1H), 7.16-7.10 (m, 1H), 7.09-7.04 (m, 1H), 6.96 (t, J=10.9Hz, 1H), 5.39-4.97 (m, 1H), 3.84-3.59 (m, 2H), 3.48 (d, J=4.9 Hz, 1H),3.09-2.86 (m, 2H), 1.25-1.19 (m, 6H); ES-LCMS m/z 332.1, 334.1 [M+H]⁺.

Step 3:4-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-1,3,5-triazin-2-amine(I-136)

To a solution of4-chloro-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-1,3,5-triazin-2-amine(90 mg, 244.12 μmol, 1 eq) in 1,4-dioxane (3 mL) and H₂O (1 mL) wasadded (5-fluoro-3-pyridyl)boronic acid (34.40 mg, 244.12 μmol, 1 eq),Cs₂CO₃ (238.62 mg, 732.37 μmol, 3 eq) and Pd(dppf)Cl₂ (17.86 mg, 24.41μmol, 0.1 eq) under N₂. The mixture was stirred at 100° C. for 3 h underN₂. The reaction mixture was filtered and concentrated to yield aresidue which was purified by preparative HPLC (HCl condition; column:Agela ASB 150*25 mm*5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %:55%-85%, 8 min) to yield4-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-1,3,5-triazin-2-amine(25.60 mg, 55.01 μmol, 22.5% yield, 100.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.80 (s, 1H), 9.24 (d, J=16.1Hz, 1H), 8.75 (s, 1H), 8.38-8.18 (m, 2H), 7.56 (dd, J=8.2, 13.8 Hz, 1H),7.29 (t, J=8.4 Hz, 1H), 7.16 (s, 1H), 7.06-6.99 (m, 1H), 6.98-6.90 (m,1H), 5.36-5.16 (m, 1H), 3.75-3.51 (m, 2H), 2.95 (s, 2H), 1.34-1.26 (m,6H); ES-LCMS m/z 393.1 [M+H]⁺.

Example 125

Synthesis of I-137a, I-137b and I-137c

Synthetic Scheme:

Step 1: 1,4,6,7-Tetrahydroindol-5-one oxime

A mixture of 1,4,6,7-tetrahydroindol-5-one (150 mg, 1.11 mmol, 1 eq),NH₂OH HCl (92.54 mg, 1.33 mmol, 1.2 eq) and NaOAc (136.56 mg, 1.66 mmol,1.5 eq) in THE (10 mL) was stirred at 60° C. for 3 h. TLC (PE/EtOAc=1/1,R_(f)=0.20) showed the starting material was consumed completely. Thereaction mixture was diluted with H₂O (20 mL) and extracted with EtOAc(20 mL×3). The organic layer was dried over Na₂SO₄, filtered andconcentrated under reduced pressure to yield1,4,6,7-tetrahydroindol-5-one oxime (160 mg, crude) as a brown gum whichwas used in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) (7.81 (br s, 1H), 6.69-6.64 (m, 1H), 6.06-5.98 (m, 1H), 3.63-3.35(m, 2H), 2.81-2.74 (m, 2H), 2.68-2.53 (m, 2H); ES-LCMS m/z 151.1 [M+H]⁺.

Step 2: 4,5,6,7-Tetrahydro-1H-indol-5-amine

A mixture of 1,4,6,7-tetrahydroindol-5-one oxime (160 mg, 1.07 mmol, 1eq) and Raney-Ni (200 mg) in MeOH (10 mL) was stirred under H₂ (15 Psi)at 10° C. for 12 h. The reaction mixture was filtered. The filtrate wasconcentrated under reduced pressure to yield4,5,6,7-tetrahydro-1H-indol-5-amine (140 mg, crude) as a brown solidwhich was used in the next step without further purification. ¹H NMR(400 MHz, CD₃OD) δ ppm 6.55 (d, J=2.4 Hz, 1H), 5.80 (d, J=2.4 Hz, 1H),3.17-3.05 (m, 1H), 2.79 (dd, J=4.8, 14.8 Hz, 1H), 2.71-2.63 (m, 2H),2.33 (dd, J=8.8, 14.8 Hz, 1H), 2.06-1.96 (m, 1H), 1.77-1.64 (m, 1H);ES-LCMS m/z 137.2 [M+H]⁺.

Step 3:2-(5-fluoro-3-pyridyl)-8-isopropyl-N-[(5S)-4,5,6,7-tetrahydro-1H-indol-5-yl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-137a) and2-(5-fluoro-3-pyridyl)-8-isopropyl-N-[(5R)-4,5,6,7-tetrahydro-1H-indol-5-yl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-137b)

To a solution of4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(120.48 mg, 330.41 μmol, 1 eq) and 4,5,6,7-tetrahydro-1H-indol-5-amine(45 mg, 330.41 μmol, 1 eq) in i-PrOH (10 mL) was added DIEA (341.63 mg,2.64 mmol, 460.41 μL, 8 eq). The mixture was stirred at 50° C. for 2 h.TLC (PE/EtOAc=3/1, R_(f)=0.5) indicated the starting material wasconsumed completely. The reaction mixture was concentrated under reducedpressure to yield a residue which was purified by flash silica gelchromatography (from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1, R_(f)=0.50)to yield product which was separated by chiral SFC (column: DAICELCHIRALCEL OJ-H (250 mm*30 mm, 5 um); mobile phase: [0.1% NH₃H₂O MeOH]; B%: 35%-35%) to yield peak 1 (SFC: Rt=3.442) and peak 2 (SFC: Rt=3.780).Peak 1 was concentrated to yield a residue which was purified bypreparative HPLC (column: Xtimate C18 150*25 mm*5 um; mobile phase:[water(0.05% HCl)-ACN]; B %: 75%-100%, 8 min), followed bylyophilization to yield an enantiomer (17.61 mg, 35.16 μmol, 10.6%yield, 100.0% purity, 3HCl, SFC: Rt=3.442, ee=100%, OR: [α]^(20.2)_(D)=−0.069 (MeOH c=0.086 g/100 mL)) as a gray solid. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 10.37 (br s, 1H), 9.36 (s, 1H), 8.87 (d, J=8.8 Hz, 1H),8.71 (d, J=2.7 Hz, 1H), 8.46-8.39 (m, 1H), 8.13 (s, 1H), 6.56 (t, J=2.4Hz, 1H), 5.77 (t, J=2.4 Hz, 1H), 4.66 (br s, 1H), 3.27-3.17 (m, 1H),2.86-2.74 (m, 3H), 2.68 (d, J=7.8 Hz, 1H), 2.12-1.97 (m, 2H), 1.36 (d,J=6.8 Hz, 6H); ES-LCMS m/z 392.2 [M+H]⁺. Peak 2 (SFC: Rt=3.780) wasconcentrated to yield a residue which was purified by preparative HPLC(column: Xtimate C18 150*25 mm*5 um; mobile phase: [water(0.05%HCl)-ACN]; B %: 75%-100%, 8 min), followed by lyophilization to yieldthe other enantiomer (16.18 mg, 32.31 μmol, 9.8% yield, 100.0% purity,3HCl, SFC: Rt=3.780, ee=99.12%, OR: [α]^(20.3) _(D)=0.066 (MeOH c=0.084g/100 mL)) as a gray solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.38 (br s,1H), 9.36 (s, 1H), 8.88 (d, J=8.6 Hz, 1H), 8.71 (d, J=2.4 Hz, 1H),8.46-8.38 (m, 1H), 8.13 (s, 1H), 6.56 (br s, 1H), 5.77 (br s, 1H), 4.66(br s, 1H), 3.21 (td, J=6.8, 13.7 Hz, 1H), 2.84-2.72 (m, 3H), 2.71-2.66(m, 1H), 2.12-1.99 (m, 2H), 1.36 (d, J=6.8 Hz, 6H); ES-LCMS m/z 392.2[M+H]⁺.

Example 126

Synthesis of I-139a, I-139b and I-139c

Synthetic Scheme:

Step 1:(3S)-6,8-difluoro-N-[2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-139a) and(3R)-6,8-difluoro-N-[2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-139b)

To a solution of 6,8-difluoro-2,3,4,9-tetrahydro-1H-carbazol-3-amine(180 mg, 509.83 μmol, 1 eq, TFA) and4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(221.40 mg, 607.18 μmol, 1.19 eq) in i-PrOH (10 mL) was added DIEA(527.13 mg, 4.08 mmol, 710.42 μL, 8 eq). The mixture was stirred at 50°C. for 2 h. TLC (PE/EtOAc=3/1, R_(f)=0.5) indicated the startingmaterial was consumed completely. The reaction mixture was concentratedunder reduced pressure to dryness to yield a residue which was purifiedby flash silica gel chromatography (from PE/EtOAc=1/0 to 3/1, TLC:PE/EtOAc=3/1, R_(f)=0.50). The desired fraction was concentrated underreduced pressure to yield a product which was separated by chiral SFC(column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 um); mobile phase: [0.1%NH₃H₂O IPA]; B %: 45%-45%, min) to yield peak 1 (SFC: Rt=1.851) and peak2 (SFC: Rt=2.254). Peak 1 was concentrated to yield a residue which waspurified by preparative HPLC (column: Agela ASB 150*25 mm*5 um; mobilephase: [water (0.05% HCl)-ACN]; B %: 75%-100%, 8 min). The desiredfraction was lyophilized to yield an enantiomer (64.24 mg, 114.88 μmol,22.5% yield, 98.4% purity, 2HCl, SFC: Rt=1.851, ee=100%, OR: [α]^(20.2)_(D)=0.206 (MeOH c=0.110 g/100 mL)) as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 11.38 (s, 1H), 9.38 (s, 1H), 9.06 (d, J=8.6 Hz, 1H), 8.69(d, J=2.2 Hz, 1H), 8.44 (d, J=10.0 Hz, 1H), 8.15 (s, 1H), 7.02 (d, J=9.0Hz, 1H), 6.86 (m, J=10.6 Hz, 1H), 4.84 (br s, 1H), 3.22 (d, J=7.1, 14.0Hz, 1H), 3.12-2.98 (m, 2H), 2.88 (d, J=15.4 Hz, 2H), 2.20 (br s, 2H),1.37 (d, J=6.8 Hz, 6H); ES-LCMS m/z 478.3 [M+H]⁺. Peak 2 wasconcentrated to yield a residue which was purified by preparative HPLC(column: Agela ASB 150*25 mm*5 um; mobile phase: [water(0.05% HCl)-ACN];B %: 75%-100%, 8 min). The desired fraction was lyophilized to yield theother enantiomer (70.3 mg, 125.02 μmol, 24.5% yield, 97.9% purity, 2HCl,SFC: Rt=2.254, ee=99.79%, OR: [α]^(20.2) _(D)=−0.239 (MeOH c=0.117 g/100mL)) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.38 (s, 1H),9.38 (s, 1H), 9.05 (d, J=8.6 Hz, 1H), 8.69 (d, J=2.9 Hz, 1H), 8.49-8.38(m, 1H), 8.15 (s, 1H), 7.02 (dd, J=1.8, 9.4 Hz, 1H), 6.91-6.80 (m, 1H),4.94-4.75 (m, 1H), 3.22 (d, J=6.9, 13.8 Hz, 1H), 3.13-2.98 (m, 2H),2.96-2.83 (m, 2H), 2.28-2.13 (m, 2H), 1.37 (d, J=6.8 Hz, 6H); ES-LCMSm/z 478.2 [M+H]⁺.

Example 127

Synthesis of I-141

Synthetic Scheme:

Step 1:8-Bromo-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine (I-141)

To a solution of8-bromo-4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine(50 mg, 152.19 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (59.01 mg,456.58 μmol, 79.53 μL, 3 eq) and 2-(1H-indol-3-yl)ethanamine (29.26 mg,182.63 μmol, 1.2 eq). The mixture was stirred at 60° C. for 2 h. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: Xtimate C18150*25 mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 55%-85%, 8min;), followed by lyophilization to yield8-bromo-2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(19.80 mg, 37.70 μmol, 24.8% yield, 100.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.81 (s, 1H), 9.34 (s, 2H), 8.76(s, 1H), 8.38 (s, 1H), 8.29 (d, J=9.8 Hz, 1H), 7.66 (d, J=7.6 Hz, 1H),7.29 (d, J=8.1 Hz, 1H), 7.20 (s, 1H), 7.04 (t, J=7.6 Hz, 1H), 7.00-6.91(m, 1H), 3.95 (t, J=7.2 Hz, 2H), 3.13 (t, J=7.2 Hz, 2H); ES-LCMS m/z452.1, 454.1 [M+H]⁺.

Example 128

Synthesis of I-143

Synthetic Scheme:

Step 1:3-[2-[[5-(5-Fluoro-3-pyridyl)-3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-1H-indol-5-ol (I-143)

To a solution of[7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-3-yl]methanol(80 mg, 287.08 μmol, 1 eq) in i-PrOH (4 mL) was added3-(2-aminoethyl)-1H-indol-5-ol (50.59 mg, 287.08 μmol, 1 eq) and DIEA(111.31 mg, 861.24 μmol, 150.01 μL, 3 eq). The mixture was stirred at90° C. for 3 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 μm;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 20%-50%, 8min), followed by lyophilization to yield3-[2-[[5-(5-fluoro-3-pyridyl)-3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-1H-indol-5-ol(3.76 mg, 8.99 μmol, 3.1% yield, 100% purity) as a white solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 8.73 (s, 1H), 8.48 (s, 1H), 8.06 (s, 1H), 7.78(d, J=9.5 Hz, 1H), 7.09-7.02 (m, 2H), 6.93 (s, 1H), 6.67 (d, J=8.8 Hz,1H), 6.05 (s, 1H), 4.80 (s, 2H), 3.86 (t, J=6.1 Hz, 2H), 3.13 (t, J=5.9Hz, 2H); ES-LCMS m/z 440.8 [M+Na]⁺.

Example 129

Synthesis of I-144a

Synthetic Scheme:

Step 1:7-Chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde

To a solution of DMF (10 mL) was added POCl₃ (5 g) dropwise at −20° C.over a period of 12 mins under N₂ atmosphere. After 1 h,7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (1 g, 3.76mmol, 1 eq) in DMF (5 mL) was added to the above solution during whichthe temperature was maintained below −20° C. The mixture was warmed to20° C. and stirred for 12 h under N₂ atmosphere. POCl₃ (12.85 g) in DMF(10 mL) was added to the above solution. The mixture was stirred at 20°C. for 12 h under N₂ atmosphere. The mixture was concentrated and water(200 mL) was added. The mixture was extracted with DCM (50 mL×3). Thecombined organic layers were washed with 5% LiCl (50 mL), dried overNa₂SO₄, filtered and concentrated to yield7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde(800 mg, 2.77 mmol, 73.5% yield, 95.7% purity) as a yellow solid whichwas used in the next step without further purification. ¹H NMR (400 MHz,CD₃OD) δ ppm 10.29 (s, 1H), 9.42 (s, 1H), 8.9-8.79 (m, 2H), 8.63 (d,J=10.0 Hz, 1H), 8.52 (s, 1H); ES-LCMS m/z 277.0 [M+H]⁺.

Step 2:[7-Chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-3-yl]methanol

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde(400 mg, 1.38 mmol, 1 eq) in THE (10 mL) was added NaBH₄ (78.52 mg, 2.08mmol, 1.5 eq) at 0° C., the mixture was stirred at 20° C. for 12 h underN₂ atmosphere. To the reaction mixture was added 0.5 N aq.HCl (0.05 mL).The mixture was concentrated under reduced pressure to yield a residuewhich was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=1/1, R_(f)=0.27) to yield[7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-3-yl]methanol(130 mg, 421.25 μmol, 30.4% yield, 90.3% purity) as a yellow solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 10.28 (s, 1H), 9.70 (d, J=2.2 Hz, 1H), 9.48(d, J=10.3 Hz, 1H), 9.33 (s, 1H), 9.15 (s, 1H), 6.10 (br s, 1H), 5.71(br s, 2H); ES-LCMS m/z 279.0 [M+H]⁺.

Step 3:(1S)-2-[[5-(5-Fluoro-3-pyridyl)-3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol (I-144a)

To a solution of[7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-3-yl]methanol(55 mg, 178.22 μmol, 1 eq) in i-PrOH (3 mL) was added(1S)-2-amino-1-(1H-indol-3-yl)ethanol (36.95 mg, 178.22 μmol, 1 eq) andDIEA (69.10 mg, 534.66 μmol, 93.13 μL, 3 eq). The mixture was stirred at90° C. for 3 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 μm;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 25%-55%, 8min), followed by lyophilization to yield(1S)-2-[[5-(5-fluoro-3-pyridyl)-3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol(40 mg, 94.55 μmol, 53.0% yield, 98.9% purity, SFC: R_(t)=4.476,ee=99.1%, [α]^(19.6) _(D)=−11.254 (MeOH, c=0.106 g/100 mL)) as a whitesolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.73 (s, 1H), 8.48 (d, J=2.2 Hz,1H), 8.07 (s, 1H), 7.88-7.81 (m, 2H), 7.29-7.21 (m, 1H), 7.18 (s, 1H),7.13-7.05 (m, 2H), 6.16 (s, 1H), 5.30 (t, J=5.6 Hz, 1H), 4.81 (s, 2H),4.07-3.95 (m, 2H); ES-LCMS m/z 419.1 [M+H]⁺.

Example 130

Synthesis of I-144b

Synthetic Scheme:

Step 1:(1R)-2-[[5-(5-Fluoro-3-pyridyl)-3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol (I-144b)

To a solution of[7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-3-yl]methanol(66.45 mg, 215.31 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (83.48 mg,645.93 μmol, 112.51 μL, 3 eq) and (1R)-2-amino-1-(1H-indol-3-yl)ethanol(53.32 mg, 279.90 μmol, 1.3 eq). The reaction mixture was stirred at 80°C. for 12 h. The reaction mixture was concentrated under reducedpressure to yield a residue which was purified by preparative HPLC twice(basic condition; column: Phenomenex Gemini 150*25 mm*10 μm; mobilephase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 25%-55%, 8 min).The desired fraction was lyophilized to yield(1R)-2-[[5-(5-fluoro-3-pyridyl)-3-(hydroxymethyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol(18.50 mg, 44.21 μmol, 20.5% yield, 100.0% purity, SFC: R_(t)=3.628 min,ee=99.4%, [α]^(19.5) _(D)=+12.650 (MeOH, c=0.106 g/100 mL)) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.74 (s, 1H), 8.48 (d, J=2.4 Hz,1H), 8.08 (s, 1H), 7.90-7.80 (m, 2H), 7.25 (d, J=8.3 Hz, 1H), 7.18 (s,1H), 7.13-7.05 (m, 2H), 6.17 (s, 1H), 5.31 (t, J=5.7 Hz, 1H), 4.81 (s,2H), 4.09-3.95 (m, 2H); ES-LCMS m/z 419.1 [M+H]⁺.

Example 131

Synthesis of I-150a

Synthetic Scheme:

Step 1:(1R)-2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol(I-150a)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 185.97 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (120.18 mg,929.87 μmol, 161.96 μL, 5 eq) and (1R)-2-amino-1-(1H-indol-3-yl)ethanol(46.06 mg, 241.77 μmol, 1.3 eq). The mixture was stirred at 90° C. for12 h. The reaction mixture was concentrated under reduced pressure toyield a residue which was purified by flash silica gel chromatography(from PE/EtOAc=1/0 to 1/2, TLC: PE/EtOAc=1/1, R_(f)=0.12) to yield(1R)-2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol(15.35 mg, 38.14 μmol, 20.5% yield, 100% purity, SFC: R_(t)=1.560,ee=99.008%, [α]^(22.1) _(D)=+6.048 (MeOH, c=0.133 g/100 mL)) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.73 (s, 1H), 8.48 (d, J=2.8 Hz,1H), 7.89 (s, 1H), 7.85-7.79 (m, 2H), 7.25 (d, J=7.0 Hz, 1H), 7.19 (s,1H), 7.12-7.05 (m, 2H), 6.11 (s, 1H), 5.30 (t, J=5.6 Hz, 1H), 4.06-3.96(m, 2H), 2.31 (s, 3H); ES-LCMS m/z 403.1 [M+H]⁺.

Example 132

Synthesis of I-150b

Synthetic Scheme:

Step 1: 2-(1H-Indol-3-yl)-2-oxo-acetyl chloride

To a solution of indole (10 g, 85.36 mmol, 1 eq) in THF (100 mL) wasadded drop-wise (COCl)₂ (11.05 g, 87.07 mmol, 7.62 mL, 1.02 eq) at 0-5°C. under N₂. The mixture was stirred at 0-5° C. for 3 h. The yellowslurry was filtered, the cake was washed with PE (10 mL×2), dried underreduced pressure to yield crude 2-(1H-indol-3-yl)-2-oxo-acetyl chloride(15 g, 72.25 mmol, 84.6% yield, 100% purity) as a yellow solid which wasused in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 13.31 (br s, 1H), 8.64-8.57 (m, 1H), 8.34-8.25 (m, 1H),7.45-7.38 (m, 1H), 7.26-7.15 (m, 2H).

Step 2: 2-(1H-Indol-3-yl)-2-oxo-acetamide

To a solution of NH₃.H₂O (42.20 g, 337.17 mmol, 46.37 mL, 28%, 10 eq) inEtOH (100 mL) was added 2-(1H-indol-3-yl)-2-oxo-acetyl chloride (7 g,33.72 mmol, 1 eq). The mixture was stirred at 0° C. for 1.5 h. Theslurry was filtered, the filter cake was washed with water (20 mL×2),dried under reduced pressure to yield 2-(1H-indol-3-yl)-2-oxo-acetamide(5.5 g, 28.00 mmol, 83.0% yield, 95.8% purity) as a white solid whichwas used in the next step without further purification. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 12.18 (br s, 1H), 8.69 (s, 1H), 8.27-8.17 (m, 1H), 8.08(br s, 1H), 7.71 (br s, 1H), 7.58-7.48 (m, 1H), 7.30-7.19 (m, 2H);ES-LCMS m/z 189.1 [M+H]⁺.

Step 3: 1H-Indole-3-carbonyl cyanide

To a solution of 2-(1H-indol-3-yl)-2-oxo-acetamide (5.5 g, 28.00 mmol, 1eq) and pyridine (6.64 g, 84.00 mmol, 6.78 mL, 3 eq) in EtOAc (100 mL)was added TFAA (8.82 g, 42.00 mmol, 5.84 mL, 1.5 eq) at 10° C. under N₂.The mixture was stirred at 10° C. for 4 h. The reaction mixture wasquenched by addition of NaHCO₃ (100 mL), extracted with EtOAc (80 mL×3).The combined organic layers were washed with 0.5N aq. HCl (20 mL), brine(20 mL), dried over Na₂SO₄, filtered and concentrated under reducedpressure to yield crude 1H-indole-3-carbonyl cyanide (3.2 g, 12.73 mmol,45.5% yield, 67.7% purity) as a white solid which was used in the nextstep without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.88(br s, 1H), 8.63 (s, 1H), 8.07-8.01 (m, 1H), 7.58 (dd, J=1.3, 7.0 Hz,1H), 7.34 (dq, J=1.3, 7.2 Hz, 2H); ES-LCMS m/z 171.1 [M+H]⁺.

Step 4: (1S)-2-Amino-1-(1H-indol-3-yl)ethanol

To a solution of 1H-indole-3-carbonyl cyanide (1 g, 3.98 mmol, 1 eq) inTHE (40 mL) was added LAH (302.00 mg, 7.96 mmol, 2 eq) at 0° C. underN₂. The mixture was stirred at 15° C. for 2 h. TLC (PE/EtOAc=3/1,R_(f)=0.18) showed the starting material was consumed completely. Themixture was diluted with THE (50 mL), cooled to 0° C., quenched by water(0.3 mL), 10% aq. NaOH (0.3 mL), water (0.9 mL) in sequence. After beingstirred for 30 min, the mixture was filtered through a celite. Thefiltrate was concentrated under reduced pressure to yield a residuewhich was purified by flash silica gel chromatography (fromEtOAc/MeOH=1/0 to 1/1, TLC: EtOAc/MeOH=10/1, R_(f)=0.18) to yieldproduct which was purified by SFC separation twice (first: DAICELCHIRALPAK IC (250 mm*30 mm, 10 μm); mobile phase: [0.1% NH₃H₂O IPA]; B%: 35%-35%. second: column: DAICEL CHIRALPAK IC (250 mm*30 mm, 5 μm);mobile phase: [0.1% NH₃H₂O IPA]; B %: 45%-45%) to yield(1R)-2-amino-1-(1H-indol-3-yl)ethanol (120 mg, 629.91 μmol, 15.8% yield,92.5% purity, ee=92.5%) as yellow oil. ¹H NMR (400 MHz, CD₃OD) δ ppm7.66 (d, J=7.8 Hz, 1H), 7.35 (d, J=8.3 Hz, 1H), 7.22 (s, 1H), 7.10 (t,J=7.2 Hz, 1H), 7.05-6.95 (m, 1H), 4.96 (t, J=6.3 Hz, 1H), 3.02 (d, J=6.3Hz, 2H); ES-LCMS m/z 159.2 [M−H₂O+H]⁺. And(1S)-2-amino-1-(1H-indol-3-yl)ethanol (125 mg, 602.96 μmol, 15.2% yield,85.0% purity, ee=98.698%) as yellow oil. ¹H NMR (400 MHz, CD₃OD) δ ppm7.67 (d, J=8.0 Hz, 1H), 7.35 (d, J=8.0 Hz, 1H), 7.23 (s, 1H), 7.11 (t,J=7.2 Hz, 1H), 7.05-6.99 (m, 1H), 4.99 (t, J=6.3 Hz, 1H), 3.08-3.02 (m,2H); ES-LCMS m/z 159.2 [M−H₂O+H]⁺.

Step 5(1S)-2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol(I-150b)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 185.97 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (72.11 mg, 557.92μmol, 97.18 μL, 3 eq) and (1S)-2-amino-1-(1H-indol-3-yl)ethanol (57.47mg, 277.22 μmol, 1.49 eq). The mixture was stirred at 90° C. for 12 h.The reaction mixture was concentrated under reduced pressure to yield aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 1/2, TLC: PE/EtOAc=1/1, R_(f)=0.18) to yield(1S)-2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol(29.26 mg, 71.26 μmol, 38.3% yield, 98.0% purity, SFC: R_(t)=2.204,ee=98.378, [α]^(21.4) _(D)=−8.227 (MeOH, c=0.10 g/100 mL)) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.72 (s, 1H), 8.48 (d, J=2.2 Hz,1H), 7.89 (s, 1H), 7.82 (d, J=5.9 Hz, 2H), 7.25 (d, J=8.3 Hz, 1H), 7.19(s, 1H), 7.14-7.02 (m, 2H), 6.10 (s, 1H), 5.30 (t, J=5.7 Hz, 1H),4.08-3.95 (m, 2H), 2.31 (s, 3H); ES-LCMS m/z 403.2 [M+H]⁺.

Example 133

Synthesis of I-152a, I-152b, I-152c

Synthetic Scheme:

Step 1:(3R)-6,8-difluoro-N-[2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-152b) and(3S)-6,8-difluoro-N-[2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-152a)

To a solution of 6,8-difluoro-2,3,4,9-tetrahydro-1H-carbazol-3-amine(180 mg, 509.83 μmol, 1 eq, TFA) and4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(221.40 mg, 607.18 μmol, 1.19 eq) in i-PrOH (10 mL) was added DIEA(527.13 mg, 4.08 mmol, 710.42 μL, 8 eq). The mixture was stirred at 50°C. for 2 h. TLC (PE/EtOAc=3/1, R_(f)=0.5) indicated the startingmaterial was consumed completely. The reaction mixture was concentratedunder reduced pressure to dryness to yield a residue which was purifiedby flash silica gel chromatography (from PE/EtOAc=1/0 to 3/1, TLC:PE/EtOAc=3/1, R_(f)=0.50). The desired fraction was concentrated underreduced pressure to yield a product which was separated by chiral SFC(column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 um); mobile phase: [0.1%NH₃H₂O IPA]; B %: 45%-45%, min) to yield peak 1 (SFC: Rt=1.851) and peak2 (SFC: Rt=2.254). Peak 1 was concentrated to yield a residue which waspurified by preparative HPLC (column: Agela ASB 150*25 mm*5 um; mobilephase: [water (0.05% HCl)-ACN]; B %: 75%-100%, 8 min). The desiredfraction was lyophilized to yield an enantiomer (64.24 mg, 114.88 μmol,22.5% yield, 98.4% purity, 2HCl, SFC: Rt=1.851, ee=100%, OR: [α]^(20.2)_(D)=0.206 (MeOH c=0.110 g/100 mL)) as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 11.38 (s, 1H), 9.38 (s, 1H), 9.06 (d, J=8.6 Hz, 1H), 8.69(d, J=2.2 Hz, 1H), 8.44 (d, J=10.0 Hz, 1H), 8.15 (s, 1H), 7.02 (d, J=9.0Hz, 1H), 6.86 (m, J=10.6 Hz, 1H), 4.84 (br s, 1H), 3.22 (d, J=7.1, 14.0Hz, 1H), 3.12-2.98 (m, 2H), 2.88 (d, J=15.4 Hz, 2H), 2.20 (br s, 2H),1.37 (d, J=6.8 Hz, 6H); ES-LCMS m/z 478.3 [M+H]⁺. Peak 2 wasconcentrated to yield a residue which was purified by preparative HPLC(column: Agela ASB 150*25 mm*5 um; mobile phase: [water(0.05% HCl)-ACN];B %: 75%-100%, 8 min). The desired fraction was lyophilized to yield theother enantiomer (70.3 mg, 125.02 μmol, 24.5% yield, 97.9% purity, 2HCl,SFC: Rt=2.254, ee=99.79%, OR: [α]^(20.2) _(D)=−0.239 (MeOH c=0.117 g/100mL)) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.38 (s, 1H),9.38 (s, 1H), 9.05 (d, J=8.6 Hz, 1H), 8.69 (d, J=2.9 Hz, 1H), 8.49-8.38(m, 1H), 8.15 (s, 1H), 7.02 (dd, J=1.8, 9.4 Hz, 1H), 6.91-6.80 (m, 1H),4.94-4.75 (m, 1H), 3.22 (d, J=6.9, 13.8 Hz, 1H), 3.13-2.98 (m, 2H),2.96-2.83 (m, 2H), 2.28-2.13 (m, 2H), 1.37 (d, J=6.8 Hz, 6H); ES-LCMSm/z 478.2 [M+H]⁺.

Example 134

Synthesis of I-154

Synthetic Scheme:

Step 1:8-Bromo-4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine

To a solution of4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine (110 mg,423.02 μmol, 1 eq) in MeCN (2 mL) and DCM (1 mL) was added NBS (82.82mg, 465.33 μmol, 1.1 eq). The mixture was stirred at 30° C. for 2 h. Thereaction mixture was concentrated under reduced pressure to yield8-bromo-4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine(120 mg, 365.27 μmol, 86.4% yield) as a yellow solid which was used inthe next step without further purification. ¹H NMR (400 MHz, CDCl₃) δppm 9.56 (s, 1H), 8.68 (d, J=2.4 Hz, 1H), 8.51 (d, J=9.3 Hz, 1H), 8.32(s, 1H); ES-LCMS m/z 327.9, 329.9 [M+H]⁺.

Step 2:(3R)—N-[8-Bromo-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-154)

A mixture of8-bromo-4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine(70 mg, 204.55 μmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(41.91 mg, 225.00 μmol, 1.1 eq) and DIEA (79.31 mg, 613.65 μmol, 106.89μL, 3 eq) in i-PrOH (3 mL) was degassed and purged with N₂ for 3 timesThe mixture was stirred at 60° C. for 3 h under N₂ atmosphere. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: Agela ASB 150*25mm*5 um; mobile phase: [water(0.05% HCl)-ACN]; B %: 63%-93%, 8 min),followed by lyophilization to yield(3R)—N-[8-bromo-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(32.48 mg, 58.92 μmol, 28.8% yield, 100.0% purity, 2HCl, OR: [α]^(22.1)_(D)=0.234 (MeOH, c=0.105 g/100 mL) as a yellow solid. ¹H NMR (400 MHz,CD₃OD) δ ppm 9.45 (s, 1H), 8.85-8.72 (m, 2H), 8.12 (s, 1H), 7.34 (d,J=7.8 Hz, 1H), 7.25 (d, J=8.1 Hz, 1H), 7.02 (t, J=7.6 Hz, 1H), 6.97-6.89(m, 1H), 4.89-4.83 (m, 1H), 3.27 (m, 1H), 3.08-2.86 (m, 3H), 2.43-2.17(m, 2H); ES-LCMS m/z 480.1, 482.1 [M+H]⁺.

Example 135

Synthesis of I-155

Synthetic Scheme:

Step 1: 4,6-Dichloro-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine

To a mixture of 2-(1H-indol-3-yl)ethanamine (666.77 mg, 4.16 mmol, 1.05eq) in dry THE (10 mL) was added NaH (237.58 mg, 5.94 mmol, 1.5 eq)under ice bath at N₂ atmosphere. After being stirred for 30 min, themixture was cooled to −60° C. and4,6-dichloro-2-methylsulfonyl-pyrimidine (900 mg, 3.96 mmol, 1 eq) indry THF (10 mL) was added dropwise and kept the temperature below −55°C. The resulting mixture was stirred for 1 h at −55° C. The reactionmixture was quenched by addition of H₂O (15 mL), diluted with H₂O (15mL) and extracted with EtOAc (20 mL×3). The combined organic layers werewashed with H₂O (15 mL×2), dried over Na₂SO₄, filtered and concentrated.The residue was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=1/3, R_(f)=0.43) to yield4,6-dichloro-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine (570 mg, 1.48mmol, 37.5% yield, 80% purity) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 7.96 (br s, 1H), 7.59 (d, J=7.8 Hz, 1H), 7.31 (d, J=8.1 Hz,1H), 7.15 (t, J=7.6 Hz, 1H), 7.11-7.05 (m, 1H), 6.99 (d, J=2.0 Hz, 1H),6.52 (s, 1H), 5.37 (br s, 1H), 3.73-3.68 (m, 2H), 3.00 (t, J=6.7 Hz,2H); ES-LCMS m/z 307.0, 309.0 [M+H]⁺.

Step 2:4-Chloro-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-2-amine

To a solution of i-PrOH (75.12 mg, 1.25 mmol, 95.70 μL, 1.2 eq) in THE(3 mL) was added NaH (62.50 mg, 1.56 mmol, 1.5 eq), the mixture wasstirred at 0° C. for 30 min. To the mixture was added a solution of4,6-dichloro-N-[2-(1H-indol-3-yl)ethyl]pyrimidin-2-amine (400 mg, 1.04mmol, 1 eq) in THF (3 mL), the mixture was stirred at 60° C. for 12 h.To the mixture was added H₂O (3 mL) and concentrated to yield a residuewhich was purified with preparative TLC (PE/EtOAc=3/1, R_(f)=0.5) toyield 4-chloro-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-2-amine(470 mg, 710.37 μmol, 68.2% yield, 50% purity) as yellow oil. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.03 (br s, 1H), 7.66-7.60 (m, 1H), 7.36 (d,J=8.1 Hz, 1H), 7.20 (t, J=7.6 Hz, 1H), 7.15-7.09 (m, 1H), 7.03 (s, 1H),6.57 (s, 1H), 5.26 (d, J=19.3 Hz, 1H), 3.78-3.69 (m, 2H), 3.05 (t, J=6.4Hz, 2H), 1.62-1.18 (m, 6H); ES-LCMS m/z 331.1 [M+H]⁺.

Step 3:N-(2-(1H-Indol-3-yl)ethyl)-4-isopropoxy-6-phenylpyrimidin-2-amine(I-155)

To a mixture of4-chloro-N-[2-(1H-indol-3-yl)ethyl]-6-isopropoxy-pyrimidin-2-amine (450mg, 680.15 μmol, 1 eq) and phenylboronic acid (124.39 mg, 1.02 mmol, 1.5eq) in 1,4-dioxane (10 mL) and H₂O (2 mL) was added Pd(dppf)Cl₂ (49.77mg, 68.01 μmol, 0.1 eq) and Cs₂CO₃ (664.81 mg, 2.04 mmol, 3 eq) under N₂atmosphere. The mixture was taken up into a microwave tube and heated at110° C. for 30 min. The mixture was concentrated to yield a residuewhich was purified by preparative HPLC (column: Agela ASB 150*25 mm*5μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 40%-70%, 8 min),followed by lyophilization to yieldN-[2-(1H-indol-3-yl)ethyl]-4-isopropoxy-6-phenyl-pyrimidin-2-amine(19.18 mg, 44.68 μmol, 6.6% yield, 95.3% purity, HCl) as a yellow solid.¹H NMR (400 MHz, CD₃OD) δ ppm 7.76 (d, J=7.3 Hz, 2H), 7.69-7.62 (m, 1H),7.58 (t, J=6.6 Hz, 3H), 7.31 (d, J=8.1 Hz, 1H), 7.12 (s, 1H), 7.07 (t, J7.3 Hz, 1H), 7.01-6.96 (m, 1H), 6.50 (s, 1H), 5.13 (td, J 6.1, 12.2 Hz,1H), 3.89 (t, J=6.6 Hz, 2H), 3.13 (t, J=6.6 Hz, 2H), 1.32 (d, J=6.1 Hz,6H); ES-LCMS m/z 373.3 [M+H]⁺.

Example 136

Synthesis of I-160a, I-160b and I-160c)

Synthetic Scheme:

Step 1. 2-[1-(Benzenesulfonyl)indol-3-yl] acetonitrile

A solution of 1-(benzenesulfonyl)-3-(bromomethyl)indole (7 g, 19.99mmol, 1 eq) in THF (50 mL) was added to a well-stirred suspension KCN(2.07 g, 31.79 mmol, 1.59 eq) in DMSO (50 mL) and THE (20 mL) dropwiseat 0° C. under N₂ atmosphere. The mixture was warmed to 25° C. andstirred for 15.5 h under N₂ atmosphere. TLC (PE/EtOAc=3/1, R_(f)=0.30)indicated one major new spot was detected and starting material wasremained. KCN (1.76 g, 27.03 mmol, 1.35 eq) was added to the mixture.The mixture was stirred for 16 h under N₂ atmosphere. Ice water (200 mL)was added to the mixture. The mixture was extracted with EtOAc (200mL×3). The combined organic layers were washed with brine (150 mL),dried over Na₂SO₄, filtered and concentrated to yield a residue whichwas purified on silica gel column chromatography (from PE/EtOAc=1/0 to10/3, TLC: PE/EtOAc=3/1, R_(f)=0.30) to yield2-[1-(benzenesulfonyl)indol-3-yl]acetonitrile (2.0 g, 5.47 mmol, 27.6%yield, 81.0% purity) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm8.02 (d, J=8.3 Hz, 1H), 7.97-7.93 (m, 2H), 7.71 (s, 1H), 7.64-7.59 (m,2H), 7.55-7.49 (m, 2H), 7.42-7.37 (m, 1H), 7.33-7.28 (m, 1H), 3.97 (d,J=1.0 Hz, 2H); ES-LCMS m/z 297.1 [M+H]⁺.

Step 2: 2-[1-(Benzenesulfonyl)indol-3-yl]propanenitrile

To a solution of 2-[1-(benzenesulfonyl)indol-3-yl]acetonitrile (1 g,2.73 mmol, 1 eq) in THE (50 mL) was added n-BuLi (2.5 M, 1.09 mL, 1 eq)dropwise at −78° C. under N₂ atmosphere. The mixture was stirred at −78°C. under N₂ atmosphere for 1 h. A solution of MeI (387.96 mg, 2.73 mmol,170.16 μL, 1 eq) in THE (5 mL) was added dropwise to the mixture. Themixture was stirred at 20° C. under N₂ atmosphere for 3 h. TLC(PE/EtOAc=3/1, R_(f)=0.38) indicated one major new spot was detected.Ice water (30 mL) was added to the mixture, the mixture was extractedwith EtOAc (50 mL×3). The combined organic layers were washed with brine(50 mL), dried over Na₂SO₄, filtered and concentrated to yield a residuewhich was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 5/1, TLC: PE/EtOAc=3/1, R_(f)=0.38) to yield2-[1-(benzenesulfonyl)indol-3-yl]propanenitrile (420 mg, 1.14 mmol,41.6% yield, 84.0% purity) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δppm 8.03 (d, J=8.3 Hz, 1H), 7.94-7.89 (m, 2H), 7.62-7.55 (m, 3H),7.52-7.46 (m, 2H), 7.42-7.37 (m, 1H), 7.34-7.29 (m, 1H), 4.13 (q, J=7.1Hz, 1H), 1.75 (d, J=7.1 Hz, 3H); ES-LCMS m/z 311.1 [M+H]⁺.

Step 3: 2-[1-(Benzenesulfonyl)indol-3-yl]propan-1-amine

To a solution of 2-[1-(benzenesulfonyl)indol-3-yl]propanenitrile (420mg, 1.14 mmol, 1 eq) in MeOH (10 mL) was added Raney-Ni (453.70 mg, 5.30mmol, 4.66 eq) and NH₃.H₂O (910.00 mg, 6.49 mmol, 1 mL, 5.71 eq). Themixture was degassed and purged with H₂ for 3 times, stirred at 25° C.for 16 h. TLC (PE/EtOAc=3/1, R_(f)=0.16) indicated one major new spotwas detected. The reaction mixture was filtered and concentrated toyield 2-[1-(benzenesulfonyl)indol-3-yl]propan-1-amine (330 mg, 818.70μmol, 72.0% yield, 78.0% purity) as white solid which was used in thenext step without further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm8.01 (d, J=8.3 Hz, 1H), 7.87 (d, J=7.5 Hz, 2H), 7.58-7.51 (m, 2H),7.48-7.40 (m, 2H), 7.38-7.30 (m, 2H), 7.26-7.21 (m, 1H), 4.93 (br s,2H), 3.08-2.93 (m, 2H), 2.92-2.86 (m, 1H), 1.34 (d, J=6.8 Hz, 3H);ES-LCMS m/z 315.1 [M+H]⁺.

Step 4: 2-(1H-Indol-3-yl)propan-1-amine

To a solution of 2-[1-(benzenesulfonyl)indol-3-yl]propan-1-amine (330mg, 818.70 μmol, 1 eq) in MeOH (10 mL) was added NaOH (327.46 mg, 8.19mmol, 10 eq). The mixture was stirred at 50° C. for 4 h under N₂atmosphere. TLC (DCM/MeOH=10/1, R_(f)=0.20) indicated one major new spotwas detected. The mixture was concentrated and water (10 mL) was added.The mixture was extracted with EtOAc (10 mL×3). The combined organiclayers were dried over Na₂SO₄, filtered and concentrated to yield2-(1H-indol-3-yl)propan-1-amine (130 mg, 746.09 μmol, 91.1% yield,crude) as white solid which was used in the next step without furtherpurification. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.05 (br s, 1H), 7.67 (d,J=8.1 Hz, 1H), 7.42-7.36 (m, 1H), 7.21 (t, J=7.5 Hz, 1H), 7.12 (t, J=7.6Hz, 1H), 7.03 (d, J=2.0 Hz, 1H), 3.18-3.10 (m, 1H), 3.05-2.92 (m, 2H),1.40-1.36 (m, 3H).

Step 5:5-(5-Fluoro-3-pyridyl)-N-[(2S)-2-(1H-indol-3-yl)propyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine&5-(5-Fluoro-3-pyridyl)-N-[(2R)-2-(1H-indol-3-yl)propyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-160a, I-160b)

To a mixture of 2-(1H-indol-3-yl)propan-1-amine (130 mg, 746.09 μmol, 1eq) and7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine(200.59 mg, 746.09 μmol, 1 eq) in i-PrOH (10 mL) was added DIEA (289.28mg, 2.24 mmol, 389.87 μL, 3 eq). The mixture was stirred at 80° C. for15 h under N₂ atmosphere and concentrated under reduced pressure toyield a residue which was purified on silica gel column chromatography(from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1, R_(f)=0.34) to yield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)propyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(300 mg, 689.22 μmol, 92.4% yield, 92.0% purity) as a yellow solid whichwere separated by SFC (column: DAICEL CHIRALCEL OJ (250 mm*30 mm, 10μm); mobile phase: [0.1% NH₃.H₂O EtOH]; B %: 40%-40%, min). The solutionwere concentrated to yield a crude products which were purified bypreparative HPLC (column: Agela ASB 150*25 mm*5 μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 48%-68%, 8 min), followed by lyophilization toyield an enantiomer (27.27 mg, 57.61 μmol, 8.4% yield, 100.0% purity,2HCl, SFC: R_(t)=3.079 min, ee=98.49%, OR: [α]^(23.9) _(D)=+48.385(MeOH, c=0.110 g/100 mL)) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δppm 8.73 (d, J=2.7 Hz, 1H), 8.39 (s, 1H), 8.09 (s, 1H), 7.54 (d, J=8.1Hz, 2H), 7.11 (d, J=8.1 Hz, 1H), 7.05 (s, 1H), 6.91 (t, J 7.6 Hz, 1H),6.76 (t, J=7.1 Hz, 1H), 5.74 (s, 1H), 4.00-3.94 (m, 2H), 3.58-3.51 (m,1H), 2.27 (s, 3H), 1.59 (d, J=7.1 Hz, 3H); ES-LCMS m/z 401.1 [M+H]⁺; andthe other enantiomer (64.75 mg, 136.78 μmol, 19.85% yield, 100% purity,2HCl, SFC: R_(t)=3.374 min, ee=98.93%, OR: [α]^(23.8) _(D)=−32.08 (MeOH,c=0.120 g/100 mL)) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.73(d, J=2.7 Hz, 1H), 8.39 (s, 1H), 8.09 (s, 1H), 7.57-7.51 (m, 2H), 7.12(d, J=8.3 Hz, 1H), 7.06 (s, 1H), 6.92 (t, J=7.6 Hz, 1H), 6.76 (t, J=7.5Hz, 1H), 5.75 (s, 1H), 4.02-3.92 (m, 2H), 3.59-3.51 (m, 1H), 2.28 (s,3H), 1.60 (d, J=7.1 Hz, 3H); ES-LCMS m/z 401.1 [M+H]⁺.

Example 137

Synthesis of I-162

Synthetic Scheme:

Step 1:(3R)—N-[2-(5-Fluoro-3-pyridyl)-9-isopropyl-purin-6-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-162)

A mixture of 6-chloro-2-(5-fluoro-3-pyridyl)-9-isopropyl-purine (60 mg,187.17 μmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (38.35mg, 205.89 μmol, 1.1 eq) and DIEA (72.57 mg, 561.51 μmol, 97.81 μL, 3eq) in i-PrOH (6 mL) was degassed and purged with N₂ for 3 times. Themixture was stirred at 80° C. for 15 h under N₂ atmosphere. The reactionmixture was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Agela ASB 150*25 mm*5 μm;mobile phase: [water (0.05% HCl)-ACN]; B %: 60%-90%, 8 min), followed bylyophilization to yield(3R)—N-[2-(5-fluoro-3-pyridyl)-9-isopropyl-purin-6-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(30.19 mg, 58.51 μmol, 31.3% yield, 99.7% purity, 2HCl, [α]^(22.3)_(D)=−22.78 (MeOH, c=0.149 g/100 mL) as a yellow solid. ¹H NMR (400 MHz,CD₃OD) δ ppm 9.57 (br s, 1H), 9.40 (s, 1H), 9.14 (J=8.0 Hz, 1H), 8.99(br s, 1H), 7.31 (J=7.5 Hz, 1H), 7.23 (d, J=8.0 Hz, 1H), 7.00 (t, J=7.5Hz, 1H), 6.94-6.87 (m, 1H), 5.23-5.12 (m, 1H), 5.01 (m, 1H), 3.28 (J=5.0Hz, 1H), 3.01-2.94 (m, 2H), 2.90 (J=6.8, 15.3 Hz, 1H), 2.42-2.32 (m,1H), 2.27 (td, J=7.0, 13.7 Hz, 1H), 1.78-1.74 (m, 6H); ES-LCMS m/z 442.1[M+H]⁺.

Example 138

Synthesis of I-163

Synthetic Scheme:

Step 1:2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (70mg, 260.36 μmol, 1 eq), 2-amino-1-(1H-indol-3-yl)ethanol (53.72 mg,260.36 μmol, 1 eq), DIEA (100.95 mg, 781.09 μmol, 136.05 μL, 3 eq) ini-PrOH (15 mL) was stirred at 80° C. for 12 h. The reaction mixture wasconcentrated to yield a residue which was purified on silica gel columnchromatography (from pure DCM to DCM/MeOH=10/1, TLC: DCM/MeOH=10/1,R_(f)=0.60) to yield2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol(80 mg, 152.08 μmol, 58.4% yield, 76.5% purity) as a brown solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.77 (s, 1H), 8.46 (s, 1H), 8.24 (s, 1H), 7.91(d, J=8.3 Hz, 1H), 7.82 (s, 1H), 7.78 (d, J=7.6 Hz, 1H), 7.35 (d, J=8.1Hz, 1H), 7.20 (d, J=2.2 Hz, 1H), 6.83-6.76 (m, 1H), 6.07 (s, 1H), 5.38(t, J=5.4 Hz, 1H), 3.92 (t, J=5.9 Hz, 2H), 2.35 (s, 3H); ES-LCMS m/z403.2 [M+H]⁺.

Step 2:2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanone(I-163)

To a solution of2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanol(80 mg, 152.08 μmol, 1 eq) in DCM (10 mL) was added MnO₂ (396.64 mg,4.56 mmol, 30 eq). The reaction mixture was stirred at 20° C. for 12 h.The reaction mixture was concentrated to yield a residue which waspurified by preparative HPLC (column: Agela DuraShell 150 mm_25 mm_5 μm;mobile phase: water (0.05% HCl)-ACN; B %: 40%-70%, 8 min) twice,followed by lyophilization to yield2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-1-(1H-indol-3-yl)ethanone(13.82 mg, 27.88 μmol, 18.3% yield, 95.5% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.95 (s, 1H), 8.76 (d, J=2.7 Hz,1H), 8.38 (d, J=3.2 Hz, 1H), 8.28-8.24 (m, 1H), 8.23-8.18 (m, 2H), 7.48(d, J=7.1 Hz, 1H), 7.29-7.20 (m, 2H), 6.87 (s, 1H), 5.24 (s, 2H), 2.40(s, 3H); ES-LCMS m/z 401.2 [M+H]⁺.

Example 139

Synthesis of I-164

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-8-methyl-pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-164)

To a solution of4-chloro-2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazine(50 mg, 182.24 μmol, 1 eq) and 2-(1H-indol-3-yl)ethanamine (32.12 mg,200.47 μmol, 1.1 eq) in i-PrOH (10 mL) was added DIEA (188.43 mg, 1.46mmol, 253.95 μL, 8 eq). The mixture was stirred at 50° C. for 3 h. TLC(PE/EtOAc=3/1, R_(f)=0.2) indicated the starting material was consumedcompletely. The reaction mixture was concentrated under reduced pressureto yield a residue which was purified by preparative HPLC (HClcondition; column: Agela DuraShell 150 mm_25 mm_5 um; mobile phase:[water (0.05% HCl)-ACN]; B %: 45%-75%, 9 min). The desired fraction waslyophilized to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-8-methyl-pyrazolo[1,5-a][1,3,5]triazin-4-amine(22.72 mg, 48.65 μmol, 26.7% yield, 98.6% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.15 (s, 1H), 8.83 (br s, 1H), 8.59(d, J=9.0 Hz, 1H), 7.95 (s, 1H), 7.68 (dd, J=2.4, 6.1 Hz, 1H), 7.20-7.12(m, 1H), 7.05-6.91 (m, 3H), 4.05 (t, J=6.7 Hz, 2H), 3.19 (t, J=6.7 Hz,2H), 2.29 (s, 3H); ES-LCMS m/z 388.2 [M+H]⁺.

Example 140

Synthesis of I-165

Synthetic Scheme:

Step 1:(3R)—N-[2-(5-Fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-165)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine (44.89 mg,172.65 μmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (39.30mg, 189.92 μmol, 1.1 eq) and DIEA (66.94 mg, 517.95 μmol, 90.22 μL, 3eq) in i-PrOH (3 mL) was degassed and purged with N₂ for 3 times. Themixture was stirred at 80° C. for 12 h under N₂ atmosphere. The reactionmixture was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Agela ASB 150*25 mm*5 um;mobile phase: [water (0.05% HCl)-ACN]; B %: 55%-85%, 8 min), followed bylyophilization to yield(3R)—N-[2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(20.39 mg, 42.52 μmol, 24.6% yield, 98.5% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.46 (s, 1H), 8.90 (d, J=9.0 Hz,1H), 8.82 (s, 1H), 8.13 (d, J=2.2 Hz, 1H), 7.35 (d, J=7.8 Hz, 1H), 7.25(d, J=8.1 Hz, 1H), 7.06-6.98 (m, 1H), 6.97-6.91 (m, 1H), 6.57 (d, J=2.0Hz, 1H), 4.89 (m, 1H), 3.31-3.26 (m, 1H), 3.11-2.88 (m, 3H), 2.42-2.34(m, 1H), 2.33-2.21 (m, 1H); ES-LCMS m/z 400.2 [M+H]⁺; Optical rotation([α]^(21.4) _(D)=0.396 (MeOH, c=0.102 g/100 mL).

Example 141

Synthesis of I-166

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-8-isopropyl-N-[2-(1H-pyrrol-3-yl)ethyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(i-166)

To a solution of4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(50 mg, 168.66 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (21.80 mg,168.66 μmol, 29.38 μL, 1 eq) and 2-(1H-pyrrol-3-yl)ethanamine (27.87 mg,252.99 μmol, 1.5 eq). The mixture was stirred at 60° C. for 3 h. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: Agela ASB 150*25mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 60%-90%, 8 min),followed by lyophilization to yield2-(5-fluoro-3-pyridyl)-8-isopropyl-N-[2-(1H-pyrrol-3-yl)ethyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(15.28 mg, 32.18 μmol, 19.1% yield, 100.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.52 (s, 1H), 9.39 (s, 1H), 8.93(t, J=5.6 Hz, 1H), 8.73 (d, J=2.9 Hz, 1H), 8.41 (d, J=8.8 Hz, 1H), 8.11(s, 1H), 6.65 (d, J=2.4 Hz, 2H), 5.98 (s, 1H), 3.84-3.75 (m, 2H),3.23-3.16 (m, 1H), 2.84 (t, J=7.3 Hz, 2H), 1.35 (d, J=6.8 Hz, 6H);ES-LCMS m/z 366.2 [M+H]⁺.

Example 142

Synthesis of I-169

Synthetic Scheme:

Step 1: Methyl 3-(5-fluoro-3-pyridyl)-2-methyl-3-oxo-propanoate

To a solution of methyl(Z)-3-(5-fluoro-3-pyridyl)-3-hydroxy-prop-2-enoate (600.00 mg, 2.83μmol, 1 eq) and MeI (401.71 mg, 2.83 μmol, 176.19 μL, 1 eq) in DMF (20mL) was added K₂CO₃ (391.14 mg, 2.83 μmol, 1 eq). The mixture wasstirred at 20° C. for 3 h. The reaction mixture was quenched by additionH₂O (50 mL), extracted with EtOAc (50 mL×3). The combined organic layerswere dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to yield a residue which was purified on silica gelcolumn chromatography (from PE/EtOAc=1/0 to 10/1, TLC: PE/EtOAc=10/1,R_(f)=0.37) to yield the product of methyl3-(5-fluoro-3-pyridyl)-2-methyl-3-oxo-propanoate (605 mg, 2.69 μmol,95.0% yield, 93.9% purity) as a yellow oil. ¹H NMR (400 MHz CDCl₃) δ ppm9.00 (d, J=1.5 Hz, 1H), 8.78-8.61 (m, 1H), 8.03-7.90 (m, 1H), 4.36 (q,J=7.1 Hz, 1H), 3.71 (s, 3H), 1.57-1.52 (m, 3H); ES-LCMS m/z 212.1[M+H]⁺.

Step 2:5-(5-Fluoro-3-pyridyl)-3,6-dimethyl-pyrazolo[1,5-a]pyrimidin-7-ol

Methyl 3-(5-fluoro-3-pyridyl)-2-methyl-3-oxo-propanoate (300 mg, 1.33μmol, 1 eq), 4-methyl-1H-pyrazol-5-amine (647.37 mg, 6.67 μmol, 5 eq)were taken up into a microwave tube in AcOH (5 mL). The sealed tube washeated at 150° C. for 1 h under microwave. The mixture was concentratedunder reduced pressure to yield5-(5-fluoro-3-pyridyl)-3,6-dimethyl-pyrazolo[1,5-a]pyrimidin-7-ol (300mg, 1.16 μmol, 87.14% yield) as a white solid which was used in the nextstep without further purification.

Step 3:7-Chloro-5-(5-fluoro-3-pyridyl)-3,6-dimethyl-pyrazolo[1,5-a]pyrimidine

A solution of5-(5-fluoro-3-pyridyl)-3,6-dimethyl-pyrazolo[1,5-a]pyrimidin-7-ol (270mg, 1.05 μmol, 1 eq) in POCl₃ (8.25 g, 53.80 μmol, 5.00 mL, 51.46 eq)was stirred at 110° C. for 3 h. The reaction mixture was pour into icewater (50 g), extracted with DCM (50 mL×3). The combined organic layerswere dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to yield a residue which was purified on silica gelcolumn chromatography (from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1,R_(f)=0.51) to yield7-chloro-5-(5-fluoro-3-pyridyl)-3,6-dimethyl-pyrazolo[1,5-a]pyrimidine(200 mg, 337.55 μmol, 32.3% yield, 46.7% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.01 (d, J=1.2 Hz, 1H), 8.68 (s, 1H), 8.61(s, 1H), 8.08 (s, 1H), 2.65 (s, 3H), 2.63 (s, 3H); ES-LCMS m/z 277.1[M+H]⁺.

Step 4:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3,6-dimethyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-169)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3,6-dimethyl-pyrazolo[1,5-a]pyrimidine(100 mg, 168.78 μmol, 1 eq) and 2-(1H-indol-3-yl)ethanamine (40.56 mg,253.17 μmol, 1.5 eq) in i-PrOH (4 mL) was added DIEA (87.25 mg, 675.11μmol, 117.59 μL, 4 eq). The mixture was stirred at 70° C. for 3 h andconcentrated to yield a residue which was purified by preparative HPLC(Phenomenex Gemini 150*25 mm*10 um; mobile phase: [water (0.05% ammoniahydroxide v/v)-ACN]; B %: 50%-80%, 8 min), followed by lyophilization toyield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3,6-dimethyl-pyrazolo[1,5-a]pyrimidin-7-amine(33.61 mg, 83.93 μmol, 49.7% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 8.49 (d, J=2.7 Hz, 1H), 8.13 (s, 1H), 7.89(s, 1H), 7.52 (d, J=7.8 Hz, 1H), 7.30-7.26 (m, 1H), 7.20 (d, J=8.1 Hz,1H), 7.07 (t, J=7.5 Hz, 1H), 6.98 (d, J=7.8 Hz, 1H), 6.71 (s, 1H),4.16-4.12 (m, 2H), 3.12-3.09 (m, 2H), 2.26 (s, 3H), 1.88 (s, 3H);ES-LCMS m/z 401.2 [M+H]⁺.

Example 143

Synthesis of I-171

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)propyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-171)

A mixture of 2-(1H-indol-3-yl)propan-1-amine (30 mg, 172.17 μmol, 1 eq)and 7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine(46.29 mg, 172.17 μmol, 1 eq) in i-PrOH (6 mL) was added DIEA (66.76 mg,516.52 μmol, 89.97 μL, 3 eq). The mixture was stirred at 80° C. for 15 hunder N₂ atmosphere. The reaction mixture was concentrated under reducedpressure to yield a residue which was purified by preparative HPLC(column: Agela ASB 150*25 mm*5 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 48%-68%, 8 min), followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)propyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(18.71 mg, 39.52 μmol, 23.0% yield, 100.0% purity, 2HCl) as an orangesolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.80 (br s, 1H), 8.44 (br s, 1H),8.09 (s, 1H), 7.64 (br s, 1H), 7.56 (d, J=8.1 Hz, 1H), 7.12 (d, J=8.1Hz, 1H), 7.05 (s, 1H), 6.92 (t, J=7.3 Hz, 1H), 6.80-6.74 (m, 1H), 5.75(s, 1H), 4.04-3.92 (m, 2H), 3.59-3.50 (m, 1H), 2.28 (s, 3H), 1.59 (d,J=7.3 Hz, 3H); ES-LCMS m/z 401.1 [M+H]⁺.

Example 144

Synthesis of I-172

Synthetic Scheme:

Step 1: 7-Methoxy-1H-indole-3-carbaldehyde

To a solution of POCl₃ (1.13 g, 7.37 mmol, 684.85 μL, 1.08 eq) in DMF(12 mL) was added 7-methoxy-1H-indole (1 g, 6.79 mmol, 884.96 μL, 1 eq)at 0° C. The mixture was stirred at 0-20° C. for 3 h under N₂atmosphere. TLC (PE/EtOAc=3/1, R_(f)=0.25) indicated one major new spotwas detected. The mixture was adjusted pH to 10-12 with 2 N NaOH. Themixture was extracted with DCM (30 mL×3). The combined organic layerswere dried over Na₂SO₄, filtered and concentrated to yield a residuewhich was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 5/2, TLC: PE/EtOAc=3/1, R_(f)=0.25) to yield7-methoxy-1H-indole-3-carbaldehyde (900 mg, 5.03 mmol, 74.1% yield,98.0% purity) as a pink solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 10.07 (s,1H), 9.18 (br s, 1H), 7.89 (d, J=7.9 Hz, 1H), 7.81 (d, J=3.1 Hz, 1H),7.24 (t, J=7.9 Hz, 1H), 6.78 (d, J=7.7 Hz, 1H), 3.98 (s, 3H); ES-LCMSm/z 176.1 [M+H]⁺.

Step 2: 7-Methoxy-3-[(E)-2-nitrovinyl]-1H-indole

A mixture of 7-methoxy-1H-indole-3-carbaldehyde (900 mg, 5.03 mmol, 1eq) and NH₄OAc (504.52 mg, 6.55 mmol, 1.3 eq) in THE (10 mL) was addedCH₃NO₂ (11.30 g, 185.12 mmol, 10 mL, 36.77 eq). The mixture was degassedand purged with N₂ for 3 times and stirred at 80° C. for 19 h under N₂atmosphere. TLC (PE/EtOAc=1/1, R_(f)=0.60) indicated one major new spotwas detected. The reaction mixture was concentrated under reducedpressure to yield a residue which was purified on silica gel columnchromatography (from PE/EtOAc=1/0 to 5/2, TLC: PE/EtOAc=1/1, R_(f)=0.60)to yield 7-methoxy-3-[(E)-2-nitrovinyl]-1H-indole (660 mg, 2.87 mmol,57.1% yield, 95.0% purity) as a orange solid. ¹H NMR (400 MHz, CDCl₃) δppm 8.92 (br s, 1H), 8.29 (d, J=13.5 Hz, 1H), 7.79 (d, J=13.5 Hz, 1H),7.63 (d, J=3.1 Hz, 1H), 7.38 (d, J=8.2 Hz, 1H), 7.26-7.24 (m, 1H), 6.80(d, J=7.7 Hz, 1H), 4.00 (s, 3H).

Step 3: 2-(7-Methoxy-1H-indol-3-yl)ethanamine

To a solution of 7-methoxy-3-[(E)-2-nitrovinyl]-1H-indole (300 mg, 1.31mmol, 1 eq) in THE (10 mL) was added LiAlH₄ (247.86 mg, 6.53 mmol, 5eq). The mixture was stirred at 80° C. for 2 h under N₂ atmosphere. TLC(PE/EtOAc=3/1, R_(f)=0.10) indicated one major new spot was detected. 2mL H₂O and 1 mL 2N NaOH was added to the mixture. The mixture wasfiltered and concentrated to yield 2-(7-methoxy-1H-indol-3-yl)ethanamine(240 mg, 1.20 mmol, 91.8% yield, 95.0% purity) as yellow solid which wasused in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 8.22 (br s, 1H), 7.23 (d, J=7.8 Hz, 1H), 7.07-7.01 (m, 2H),6.66 (d, J=7.6 Hz, 1H), 3.97 (s, 3H), 3.06-3.01 (m, 2H), 2.93-2.87 (m,2H).

Step 4:5-(5-Fluoro-3-pyridyl)-N-[2-(7-methoxy-1H-indol-3-yl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (230mg, 855.48 μmol, 1 eq), 2-(7-methoxy-1H-indol-3-yl)ethanamine (217.00mg, 1.03 mmol, 1.2 eq) and DIEA (331.70 mg, 2.57 mmol, 447.03 μL, 3 eq)in i-PrOH (10 mL) was degassed and purged with N₂ for 3 times. Themixture was stirred at 80° C. for 16 h under N₂ atmosphere. The reactionmixture was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Agela DuraShell 150 mm_25 mm_5um; mobile phase: [water (0.05% HCl)-ACN]; B %: 35%-65%, 9 min),followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-N-[2-(7-methoxy-1H-indol-3-yl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(240 mg, 490.42 μmol, 57.3% yield, 100.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.72 (d, J=2.7 Hz, 1H), 8.30 (s,1H), 8.10 (s, 1H), 7.43 (td, J=2.3, 9.0 Hz, 1H), 7.15 (d, J=7.8 Hz, 1H),6.87 (s, 1H), 6.80 (t, J=7.8 Hz, 1H), 6.48 (d, J=7.6 Hz, 1H), 5.62 (s,1H), 4.04-3.98 (m, 2H), 3.81 (s, 3H), 3.21-3.16 (m, 2H), 2.29 (s, 3H);ES-LCMS m/z 417.1 [M+H]⁺.

Step 5:3-[2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-1H-indol-7-ol(I-172)

To a solution of5-(5-fluoro-3-pyridyl)-N-[2-(7-methoxy-1H-indol-3-yl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(200 mg, 480.25 μmol, 1 eq) in DCM (10 mL) was added BBr₃ (240.63 mg,960.50 μmol, 92.55 μL, 2 eq) at 0° C. The mixture was stirred at 0-20°C. for 0.5 h under N₂ atmosphere. 1 mL MeOH was added to the mixture.The mixture was concentrated under reduced pressure to yield a residuewhich was purified by preparative HPLC (column: Agela ASB 150*25 mm*5μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 35%-65%, 8 min) followedby lyophilization to yield3-[2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-1H-indol-7-ol(22.74 mg, 45.54 μmol, 9.5% yield, 95.2% purity, 2HCl) as a orangesolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.71 (d, J=2.7 Hz, 1H), 8.35 (s,1H), 8.10 (s, 1H), 7.43 (J=9.0 Hz, 1H), 7.04 (d, J=7.8 Hz, 1H), 6.87 (s,1H), 6.69 (td, J=3.8, 7.9 Hz, 1H), 6.36 (d, J=7.6 Hz, 1H), 5.65 (s, 1H),4.04-3.98 (m, 2H), 3.21-3.15 (m, 2H), 2.29 (s, 3H); ES-LCMS m/z 403.1[M+H]⁺.

Example 145

Synthesis of I-175

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N3,N3-dimethyl-N7-[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]pyrazolo[1,5-a]pyrimidine-3,7-diamine(I-175)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-N,N-dimethyl-pyrazolo[1,5-a]pyrimidin-3-amine(50 mg, 171.40 μmol, 1 eq) in i-PrOH (5 mL) was added DIEA (66.46 mg,514.21 μmol, 89.56 μL, 3 eq) and(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (35.12 mg, 188.54 μmol, 1.1eq). The mixture was stirred at 90° C. for 12 h under N₂ atmosphere. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: Agela DuraShell150 mm_25 mm_5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 30%-60%,8 min), followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-N3,N3-dimethyl-N7-[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]pyrazolo[1,5-a]pyrimidine-3,7-diamine(22.71 mg, 43.35 μmol, 25.3% yield, 98.2% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.82 (s, 1H), 9.41 (s, 1H), 8.72(s, 1H), 8.66-8.55 (m, 2H), 8.49 (br s, 1H), 7.36 (d, J=7.6 Hz, 1H),7.31-7.25 (m, 2H), 7.04-6.98 (m, 1H), 6.96-6.91 (m, 1H), 4.56 (br s,1H), 3.37 (s, 6H), 3.18-3.01 (m, 2H), 2.99-2.82 (m, 2H), 2.28-2.09 (m,2H); ES-LCMS m/z 442.3 [M+H]⁺.

Example 146

Synthesis of I-177

Synthetic Scheme:

Step 1:2-[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]propan-2-ol(I-177)

To MeLi (1.3 M, 5 mL, 57.13 eq) was added a solution of1-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone(50 mg, 113.77 μmol, 1 eq) in THE (2 mL) dropwise under N₂ atmosphere at−20° C. The mixture was stirred under N₂ atmosphere at −20° C. for 0.5h. The reaction was quenched with water (10 mL) and extracted with EtOAc(50 mL×3). The organic layer was dried over Na₂SO₄, filtered andconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Agela Durashell C18 150*30 5 u;mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %: 40%-60%, 9 min). Thedesired fraction was lyophilized to yield2-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]propan-2-ol(13.48 mg, 31.31 μmol, 27.5% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 8.69 (s, 1H), 8.43 (d, J=2.8 Hz, 1H), 7.98(s, 1H), 7.74 (td, J=2.0, 9.6 Hz, 1H), 7.68 (dd, J=2.8, 6.0 Hz, 1H),7.24-7.16 (m, 1H), 7.09-7.02 (m, 2H), 6.98 (s, 1H), 6.00 (s, 1H), 3.86(t, J=6.0 Hz, 2H), 3.18 (t, J=6.0 Hz, 2H), 1.72 (s, 6H); ES-LCMS m/z431.1 [M+H]⁺.

Example 147

Synthesis of I-179

Synthetic Scheme:

Step 1: 7-Methoxy-1H-indole-3-carbaldehyde

To a solution of POCl₃ (1.13 g, 7.37 mmol, 684.85 μL, 1.08 eq) in DMF(12 mL) was added 7-methoxy-1H-indole (1 g, 6.79 mmol, 884.96 μL, 1 eq)at 0° C. The mixture was stirred at 0-20° C. for 3 h under N₂atmosphere. TLC (PE/EtOAc=3/1, R_(f)=0.25) indicated one major new spotwas detected. The mixture was adjusted pH to 10-12 with 2 N NaOH. Themixture was extracted with DCM (30 mL×3). The combined organic layerswere dried over Na₂SO₄, filtered and concentrated to yield the residuewhich was purified on silica gel column chromatography (fromPE/EtOAc=I/O to 5/2, TLC: PE/EtOAc=3/1, R_(f)=0.25) to yield7-methoxy-1H-indole-3-carbaldehyde (900 mg, 5.03 mmol, 74.1% yield,98.0% purity) as a pink solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 10.07 (s,1H), 9.18 (br s, 1H), 7.89 (d, J=7.9 Hz, 1H), 7.81 (d, J=3.1 Hz, 1H),7.24 (t, J=7.9 Hz, 1H), 6.78 (d, J=7.7 Hz, 1H), 3.98 (s, 3H); ES-LCMSm/z 176.1 [M+H]⁺.

Step 2: 7-Methoxy-3-[(E)-2-nitrovinyl]-1H-indole

A mixture of 7-methoxy-1H-indole-3-carbaldehyde (900 mg, 5.03 mmol, 1eq) and NH₄OAc (504.52 mg, 6.55 mmol, 1.3 eq) in THE (10 mL) was addedCH₃NO₂ (11.30 g, 185.12 mmol, 10 mL, 36.77 eq). The mixture was degassedand purged with N₂ for 3 times and stirred at 80° C. for 19 h under N₂atmosphere. TLC (PE/EtOAc=1/1, R_(f)=0.60) indicated one major new spotwas detected. The reaction mixture was concentrated under reducedpressure to yield a residue which was purified on silica gel columnchromatography (from PE/EtOAc=1/0 to 5/2, TLC: PE/EtOAc=1/1, R_(f)=0.60)to yield 7-methoxy-3-[(E)-2-nitrovinyl]-1H-indole (660 mg, 2.87 mmol,57.1% yield, 95.0% purity) as a orange solid. ¹H NMR (400 MHz, CDCl₃) δppm 8.92 (br s, 1H), 8.29 (d, J=13.5 Hz, 1H), 7.79 (d, J=13.5 Hz, 1H),7.63 (d, J=3.1 Hz, 1H), 7.38 (d, J=8.2 Hz, 1H), 7.26-7.24 (m, 1H), 6.80(d, J=7.7 Hz, 1H), 4.00 (s, 3H).

Step 3: 2-(7-Methoxy-1H-indol-3-yl)ethanamine

To a solution of 7-methoxy-3-[(E)-2-nitrovinyl]-1H-indole (300 mg, 1.31mmol, 1 eq) in THE (10 mL) was added LiAlH₄ (247.86 mg, 6.53 mmol, 5eq). The mixture was stirred at 80° C. for 2 h under N₂ atmosphere. TLC(PE/EtOAc=3/1, R_(f)=0.10) indicated one major new spot was detected.H₂O (2 mL) was added, followed by 2N NaOH (1 mL) to the mixture. Themixture was filtered and concentrated to yield2-(7-methoxy-1H-indol-3-yl)ethanamine (240 mg, 1.20 mmol, 91.8% yield,95.0% purity) as yellow solid which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.22 (br s, 1H),7.23 (d, J=7.8 Hz, 1H), 7.07-7.01 (m, 2H), 6.66 (d, J=7.6 Hz, 1H), 3.97(s, 3H), 3.06-3.01 (m, 2H), 2.93-2.87 (m, 2H).

Step 4:5-(5-Fluoro-3-pyridyl)-N-[2-(7-methoxy-1H-indol-3-yl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-179)

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (230mg, 855.48 μmol, 1 eq), 2-(7-methoxy-1H-indol-3-yl)ethanamine (217.00mg, 1.03 mmol, 1.2 eq) and DIEA (331.70 mg, 2.57 mmol, 447.03 μL, 3 eq)in i-PrOH (10 mL) was degassed and purged with N₂ for 3 times. Themixture was stirred at 80° C. for 16 h under N₂ atmosphere. The reactionmixture was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Agela DuraShell 150 mm_25 mm_5μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 35%-65%, 9 min),followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-N-[2-(7-methoxy-1H-indol-3-yl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(240 mg, 490.42 μmol, 57.3% yield, 100.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.72 (d, J=2.7 Hz, 1H), 8.30 (s,1H), 8.10 (s, 1H), 7.43 (td, J=2.3, 9.0 Hz, 1H), 7.15 (d, J=7.8 Hz, 1H),6.87 (s, 1H), 6.80 (t, J=7.8 Hz, 1H), 6.48 (d, J=7.6 Hz, 1H), 5.62 (s,1H), 4.04-3.98 (m, 2H), 3.81 (s, 3H), 3.21-3.16 (m, 2H), 2.29 (s, 3H);ES-LCMS m/z 417.1 [M+H]⁺.

Example 148

Synthesis of I-180

Synthetic Scheme:

Step 1: 2-(1H-Pyrazol-3-yl)acetonitrile

To a solution of 3-(chloromethyl)-1H-pyrazole (350 mg, 2.29 mmol, 1 eq,HCl) in CH₃CN (5 mL) was added KCN (300 mg, 4.61 mmol, 197.37 μL, 2.01eq) and H₂O (1 mL). The mixture was stirred at 50° C. for 12 h. TLC(DCM/MeOH=10/1, R_(f)=0.4) indicated starting material was consumedcompletely and one new spot formed. The reaction mixture was dilutedwith H₂O (10 mL) and extracted with EtOAc (20 mL×3). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated underreduced pressure to yield 2-(1H-pyrazol-3-yl)acetonitrile (150 mg, 1.33mmol, 58.2% yield, 95.0% purity) as a yellow oil which was used in thenext step without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm12.84 (s, 1H), 7.73 (s, 1H), 6.23 (s, 1H), 3.95 (s, 2H); ES-LCMS: Nocorrect mass was found.

Step 2: 2-(1H-Pyrazol-3-yl)ethanamine

To a solution of 2-(1H-pyrazol-3-yl)acetonitrile (150 mg, 1.33 mmol, 1eq) in MeOH (10 mL) was added Raney-Ni (600 mg, 7.00 mmol, 5.26 eq)under Ar atmosphere. The suspension was degassed and purged with H₂ for3 times. The mixture was stirred under H₂ (15 psi) at 20° C. for 12 h.TLC (DCM/MeOH=10/1, R_(f)=0.4) indicated starting material was consumedcompletely. The reaction mixture was filtered and concentrated underreduced pressure to yield 2-(1H-pyrazol-3-yl)ethanamine (100 mg, 899.73μmol, 67.6% yield, crude) as a brown solid which was used in the nextstep without further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.64(s, 1H), 7.42-7.35 (m, 1H), 5.98 (s, 1H), 3.10-2.94 (m, 2H), 2.82 (d,J=17.6 Hz, 2H); ES-LCMS: No correct mass was found.

Step 3:5-(5-Fluoro-3-pyridyl)-3-methyl-N-[2-(1H-pyrazol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-180)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (75mg, 279.53 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (180.64 mg, 1.40mmol, 243.45 μL, 5 eq) and 2-(1H-pyrazol-3-yl)ethanamine (62.14 mg,559.07 μmol, 2 eq). The mixture was stirred at 80° C. for 12 h. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: PhenomenexGemini 150*25 mm*10 um; mobile phase: [water (0.05% HCl)-ACN]; B %:15%-45%, 9 min), followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-3-methyl-N-[2-(1H-pyrazol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(21.84 mg, 48.89 μmol, 17.5% yield, 100.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.07 (s, 1H), 8.88 (d, J=2.4 Hz,1H), 8.45 (td, J=2.3, 8.9 Hz, 1H), 8.21 (d, J=2.7 Hz, 1H), 8.16 (s, 1H),7.01 (s, 1H), 6.80 (d, J=2.7 Hz, 1H), 4.16 (t, J=7.0 Hz, 2H), 3.37 (t,J=7.0 Hz, 2H), 2.40 (s, 3H); ES-LCMS m/z 338.1 [M+H]⁺.

Example 149

Synthesis of I-186

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-8-methyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-186)

To a solution of4-chloro-2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazine(50 mg, 189.64 μmol, 1 eq) in i-PrOH (5 mL) was added DIEA (73.53 mg,568.92 μmol, 99.09 μL, 3 eq) and2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethanamine (40 mg, 248.13 μmol, 1.31eq). The mixture was stirred at 80° C. for 12 h. The reaction mixturewas concentrated under reduced pressure to give the residue which waspurified by preparative HPLC (HCl condition; column: Phenomenex Gemini150*25 mm*10 um; mobile phase: [water(0.05% HCl)-ACN]; B %: 10%-40%, 9min). The desired fraction was lyophilized to yield2-(5-fluoro-3-pyridyl)-8-methyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(30.06 mg, 60.39 μmol, 31.8% yield, 100.0% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.37 (d, J=1.3 Hz, 1H), 8.92 (d,J=2.3 Hz, 1H), 8.82 (d, J=8.8 Hz, 1H), 8.48 (d, J=5.8 Hz, 1H), 8.40 (d,J=8.3 Hz, 1H), 7.98 (d, J=10.5 Hz, 2H), 7.55 (dd, J=5.9, 8.2 Hz, 1H),4.19 (t, J=6.3 Hz, 2H), 3.38 (t, J=6.4 Hz, 2H), 2.29 (s, 3H); ES-LCMSm/z 389.1 [M+H]⁺.

Example 150

Synthesis of I-187b

Synthetic Scheme:

Step 1:(2S)-2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-3-(1H-indol-3-yl)propanoicacid

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (100mg, 380.71 μmol, 1 eq), (2S)-2-amino-3-(1H-indol-3-yl)propanoic acid(77.75 mg, 380.71 μmol, 1 eq), DIEA (147.61 mg, 1.14 mmol, 198.93 μL, 3eq) in i-PrOH (10 mL) was stirred at 80° C. for 12 h. The reactionmixture was concentrated to yield(2S)-2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-3-(1H-indol-3-yl)propanoicacid (160 mg, 371.72 μmol, 97.6% yield, 100.0% purity) as brown oilwhich was used in the next step without further purification. ¹H NMR(400 MHz, DMSO-d₆) δ ppm 10.69 (s, 1H), 8.91 (s, 1H), 8.57 (d, J=2.7 Hz,1H), 7.96 (d, J=10.0 Hz, 1H), 7.88 (s, 1H), 7.59-7.50 (m, 2H), 7.19 (d,J=8.1 Hz, 1H), 7.02-6.94 (m, 2H), 6.91-6.85 (m, 1H), 6.29 (s, 1H), 4.62(s, 1H), 3.58-3.54 (m, 1H), 2.94 (s, 2H), 2.21 (s, 3H); ES-LCMS m/z431.2[M+H]⁺.

Step 2:(2S)-2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-3-(1H-indol-3-yl)-N,N-dimethyl-propanamide(I-187b)

To a solution of(2S)-2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-3-(1H-indol-3-yl)propanoicacid (80 mg, 183.26 μmol, 1 eq) and N-methylmethanamine; hydrochloride(29.89 mg, 366.52 μmol, 2.0 eq) in DMF (20 mL) was added HATU (139.36mg, 366.52 μmol, 2.0 eq) and Et₃N (92.72 mg, 916.30 μmol, 127.54 μL, 5.0eq). The mixture was stirred at 25° C. for 12 h. The reaction mixturewas concentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 μm;mobile phase: [water(0.05% HCl)-ACN]; B %: 30%-60%, 9 min), followed bylyophilization to yield(2S)-2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]-3-(1H-indol-3-yl)-N,N-dimethyl-propanamide(17.77 mg, 32.74 μmol, 17.9% yield, 97.7% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.73 (d, J=2.3 Hz, 1H), 8.39 (s,1H), 8.11 (s, 1H), 7.57 (d, J=8.3 Hz, 1H), 7.45 (d, J=8.0 Hz, 1H), 7.07(d, J=8.0 Hz, 1H), 7.01 (s, 1H), 6.91 (t, J=7.4 Hz, 1H), 6.84-6.77 (m,1H), 5.83 (s, 1H), 5.40 (d, J=8.5 Hz, 1H), 3.52 (dd, J=3.0, 14.6 Hz,1H), 3.31 (s, 3H), 3.13-3.07 (m, 1H), 3.04 (s, 3H), 2.27 (s, 3H);ES-LCMS m/z 458.3 [M+H]⁺.

Example 151

Synthesis of I-192

Synthetic Scheme:

Step 1:3-Bromo-5-(5-fluoro-3-pyridyl)-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-192)

To a solution of3-bromo-7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (100mg, 305.31 μmol, 1 eq) in i-PrOH (5 mL) was added2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethanamine (50 mg, 310.17 μmol, 1.02eq) and DIEA (118.38 mg, 915.92 μmol, 159.54 μL, 3 eq). The mixture wasstirred at 60° C. for 12 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 μm;mobile phase: [water(0.05% HCl)-ACN]; B %: 25%-55%, 9 min), followed bylyophilization to yield3-bromo-5-(5-fluoro-3-pyridyl)-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(37.41 mg, 65.74 μmol, 21.5% yield, 98.7% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.11 (s, 1H), 8.72 (d, J=2.5 Hz,1H), 8.50-8.47 (m, 2H), 8.46 (s, 1H), 8.12 (s, 1H), 8.05 (s, 1H), 7.62(dd, J=5.9, 8.2 Hz, 1H), 6.69 (s, 1H), 4.03 (t, J=6.7 Hz, 2H), 3.39 (t,J=6.8 Hz, 2H); ES-LCMS m/z 452.0, 454.0 [M+H]⁺.

Example 152

Synthesis of I-195

Synthetic Scheme:

Step 1: tert-Butyl N-[2-[4-(methanesulfonamido)phenyl]ethyl]carbamate

To a solution of tert-butyl N-[2-(4-aminophenyl)ethyl]carbamate (150 mg,634.76 μmol, 1 eq) in DCM (10 mL) was added MsCl (327.20 mg, 2.86 mmol,221.08 μL, 4.5 eq) and TEA (321.16 mg, 3.17 mmol, 441.76 μL, 5 eq). Themixture was stirred at 20° C. for 20 h. The reaction mixture wasquenched by addition of saturated NaHCO₃ (50 mL), extracted with EtOAc(30 mL×3). The combined organic layers were washed with brine (10 mL),dried over Na₂SO₄, filtered and concentrated under reduced pressure toyield a residue which was purified by flash silica gel chromatography(from PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=1/1, R_(f)=0.37) to yieldtert-butyl N-[2-[4-(methanesulfonamido)phenyl]ethyl]carbamate (190 mg,186.13 μmol, 29.3% yield, 30.8% purity) as colorless oil. ¹H NMR (400MHz, CDCl₃) δ ppm 7.25-7.11 (m, 4H), 6.45 (br s, 1H), 4.58-4.49 (m, 1H),3.37 (d, J=6.8 Hz, 2H), 3.01 (s, 3H), 2.79 (t, J=7.0 Hz, 2H), 1.44 (s,9H); ES-LCMS m/z 337.1 [M+Na]⁺.

Step 2: N-[4-(2-Aminoethyl)phenyl]methanesulfonamide

To a solution of tert-butylN-[2-[4-(methanesulfonamido)phenyl]ethyl]carbamate (190 mg, 186.13 μmol,1 eq) in DCM (3 mL) was added TFA (212.23 mg, 1.86 mmol, 137.81 μL, 10eq). The mixture was stirred at 20° C. for 20 min. TLC (PE/EtOAc=1/1,R_(f)=0.06) showed that the starting material was consumed. The reactionmixture was concentrated under reduced pressure to yield a crudeN-[4-(2-aminoethyl)phenyl]methanesulfonamide (61 mg, crude, TFA) asyellow oil which was used in the next step without further purification.

Step 3:N-[4-[2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenyl]methanesulfonamide(I-195)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine(48.80 mg, 185.80 μmol, 1 eq) in i-PrOH (10 mL) was added DIEA (120.07mg, 929.01 μmol, 161.81 μL, 5 eq) andN-[4-(2-aminoethyl)phenyl]methanesulfonamide (61 mg, 185.80 μmol, 1 eq,TFA). The mixture was stirred at 60° C. for 12 h. The reaction mixturewas concentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Phenomenex Gemini C18 250*50 mm*10μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 25%-55%, 9 min),followed by lyophilization to yieldN-[4-[2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenyl]methanesulfonamide(50.35 mg, 95.81 μmol, 51.5% yield, 97.7% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.89 (s, 1H), 8.85 (d, J=2.3 Hz,1H), 8.27-8.22 (m, 1H), 8.14 (s, 1H), 7.26 (d, J=8.3 Hz, 2H), 7.08 (d,J=8.5 Hz, 2H), 6.53 (s, 1H), 4.01 (t, J=6.7 Hz, 2H), 3.08 (t, J=6.7 Hz,2H), 2.87 (s, 3H), 2.36 (s, 3H); ES-LCMS m/z 441.0 [M+H]⁺.

Example 153

Synthesis of I-196

Synthetic Scheme:

Step 1: 3-[(E)-2-Nitrovinyl]-1H-pyrrole

To a solution of 1H-pyrrole-3-carbaldehyde (1 g, 10.52 mmol, 1 eq) inCH₃NO₂ (10 mL) and THE (10 mL) was added NH₄OAc (891.60 mg, 11.57 mmol,1.1 eq). The mixture was stirred at 80° C. for 12 h. TLC (TLC:PE/EtOAc=3/1, R_(f)=0.45) indicated no starting material was remainedand one major new spot with lower polarity was detected. The reactionmixture was concentrated under reduced pressure to yield a residue whichwas purified on silica gel column chromatography (from PE/EtOAc=1/0 to10/3, TLC: PE/EtOAc=3/1, R_(f)=0.45) to yield the product3-[(E)-2-nitrovinyl]-1H-pyrrole (1.2 g, 7.82 mmol, 74.4% yield, 90.0%purity) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.64 (br s,1H), 8.03 (d, J=13.4 Hz, 1H), 7.44 (d, J=13.3 Hz, 1H), 7.22 (td, J=1.6,2.8 Hz, 1H), 6.88 (q, J=2.4 Hz, 1H), 6.49-6.43 (m, 1H).

Step 2: 2-(1H-Pyrrol-3-yl)ethanamine

To a solution of 3-[(E)-2-nitrovinyl]-1H-pyrrole (300 mg, 1.95 mmol, 1eq) in THF (20 mL) was added LiAlH₄ (370.96 mg, 9.77 mmol, 5.0 eq) underice-baths. The mixture was stirred at 80° C. for 2 h. TLC (TLC:PE/EtOAc=3/1, R_(f)=0.56) indicated no starting material was remainedand one major new spot with larger polarity was detected. The reactionmixture was quenched by addition H₂O (1.5 mL), 1N NaOH (1.5 mL) and H₂O(1.5 mL) under ice-baths, filtered and concentrated under reducedpressure to yield the crude product 2-(1H-pyrrol-3-yl)ethanamine (180mg, 1.63 mmol, 83.6% yield, crude purity) as brown oil which was used inthe next step without further purification. ¹H NMR (400 MHz, CDCl₃) δppm 8.12 (br s, 1H), 6.80-6.72 (m, 1H), 6.63 (d, J=1.3 Hz, 1H),6.14-6.08 (m, 1H), 2.93-2.85 (m, 2H), 2.63 (t, J=6.7 Hz, 2H).

Step 3:5-(5-Fluoro-3-pyridyl)-3-methyl-N-[2-(1H-pyrrol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-196)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 190.35 μmol, 1 eq) and 2-(1H-pyrrol-3-yl)ethanamine (41.94 mg,380.71 μmol, 2.0 eq) in i-PrOH (10 mL) was added DIEA (73.81 mg, 571.06μmol, 99.47 μL, 3.0 eq). The mixture was stirred at 70° C. for 3 h. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: Agela ASB 150*25mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 30%-60%, 9 min;) toyield the residue which was purified by preparative HPLC (column:Phenomenex Gemini 150*25 mm*10 um; mobile phase: [water (0.05% ammoniahydroxide v/v)-ACN]; B %: 35%-65%, 10 min), followed by lyophilizationto yield5-(5-fluoro-3-pyridyl)-3-methyl-N-[2-(1H-pyrrol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(15.72 mg, 46.73 μmol, 24.6% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 9.03 (s, 1H), 8.53 (d, J=2.7 Hz, 1H), 8.19(d, J=9.5 Hz, 2H), 7.87 (s, 1H), 6.80 (d, J=2.4 Hz, 1H), 6.72 (s, 1H),6.48 (br s, 1H), 6.27 (s, 1H), 6.19 (br s, 1H), 3.69 (q, J=6.4 Hz, 2H),2.99 (t, J=6.8 Hz, 2H), 2.39 (s, 3H); ES-LCMS m/z 337.2 [M+H]⁺.

Example 154

Synthesis of I-197

Synthetic Scheme:

Step 1:[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]methanol(I-197)

To a solution of5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (50 mg, 110.71 μmol, 1 eq) in THE (8 mL) was added LAH (50 mg, 1.32mmol, 11.90 eq) at 0° C. The mixture was stirred at 80° C. for 12 hunder N₂ atmosphere. The reaction mixture was diluted with THE (40 mL),quenched by addition of water (0.5 mL), 10% NaOH (0.5 mL) and water (1.5mL) in sequence at 0° C. After being stirred for 30 min, the mixture wasfiltered through celite. The filtrate was concentrated under reducedpressure to yield a residue which was purified by preparative HPLC(column: Xbridge 150*30 mm*10 μm; mobile phase: [water (0.05% ammoniahydroxide v/v)-ACN]; B %: 25%-55%, 8 min), followed by lyophilization toyield[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]methanol(6 mg, 14.91 μmol, 13.5% yield, 100% purity) as a yellow solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 8.65 (s, 1H), 8.47 (d, J=2.7 Hz, 1H), 8.06 (s,1H), 7.74 (d, J=9.8 Hz, 1H), 7.71-7.66 (m, 1H), 7.24-7.18 (m, 1H),7.12-7.02 (m, 2H), 6.98 (s, 1H), 6.00 (s, 1H), 4.80 (s, 2H), 3.88 (t,J=6.2 Hz, 2H), 3.20 (t, J=6.1 Hz, 2H); ES-LCMS m/z 403.2 [M+H]⁺.

Example 155

Synthesis of I-198a, I-198b and I-198c

Synthetic Scheme:

Step 1:(3R)—N-[3-bromo-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine

A mixture of3-bromo-7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (440mg, 1.34 mmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(274.54 mg, 1.47 mmol, 1.1 eq) and DIEA (519.56 mg, 4.02 mmol, 700.21μL, 3 eq) in i-PrOH (10 mL) was stirred at 80° C. for 12 h. The reactionmixture was concentrated to yield the residue which was purified onsilica gel column chromatography (from PE/EtOAc=1/0 to 2/1, TLC:PE/EtOAc=2/1, R_(f)=0.25) to yield(3R)—N-[3-bromo-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(520 mg, 1.06 mmol, 79.2% yield, 97.4% purity) as a yellow solid. ¹H NMR(400 MHz, DMSO-d₆) δ ppm 10.76 (s, 1H), 9.30 (s, 1H), 8.65 (d, J=2.9 Hz,1H), 8.46 (td, J=2.2, 10.4 Hz, 1H), 8.32-8.24 (m, 2H), 7.33 (d, J=7.5Hz, 1H), 7.24 (d, J=7.9 Hz, 1H), 7.15 (s, 1H), 7.02-6.86 (m, 2H), 4.49(td, J=4.8, 9.4 Hz, 1H), 3.14-2.98 (m, 2H), 2.94-2.79 (m, 2H), 2.23-2.10(m, 2H); ES-LCMS m/z 477.1, 479.1 [M+H]⁺.

Step 2: tert-Butyl(3R)-3-[[3-bromo-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

A mixture of(3R)—N-[3-bromo-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(500 mg, 1.02 mmol, 1 eq), Boc₂O (556.67 mg, 2.55 mmol, 585.97 μL, 2.5eq) and DMAP (373.93 mg, 3.06 mmol, 3 eq) in 1,4-dioxane (10 mL) wasstirred at 110° C. for 5 h. The reaction mixture was concentrated toyield a residue which was purified by flash silica gel chromatography(from pure PE to PE/EtOAc=3/1, TLC: PE/EtOAc=3/1, R_(f)=0.7) to yieldtert-butyl(3R)-3-[[3-bromo-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(600 mg, 664.15 umol, 65.1% yield, 75.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.06 (s, 1H), 8.60 (d, J=3.0 Hz, 1H),8.26-8.18 (m, 2H), 8.06 (d, J=8.0 Hz, 1H), 7.29 (s, 1H), 7.25-7.13 (m,3H), 4.86-4.76 (m, 1H), 3.26-3.16 (m, 2H), 2.66 (d, J=10.5 Hz, 1H), 2.36(m, 1H), 2.10-1.92 (m, 2H), 1.64 (s, 9H), 1.34 (s, 9H); ES-LCMS m/z677.2, 679.2 [M+H]⁺.

Step 3: tert-Butyl(3R)-3-[[3-acetyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

tert-Butyl(3R)-3-[[3-bromo-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(550 mg, 608.80 μmol, 1 eq), tributyl(1-ethoxyvinyl)stannane (1.02 g,2.82 mmol, 953.27 μL, 4.64 eq) and Pd(dppf)Cl₂ (89.09 mg, 121.76 μmol,0.2 eq) in toluene (8 mL) were taken up into a microwave tube and thenpurged with N₂ for 1 min. The sealed tube was heated at 120° C. for 5 hunder microwave (1 bar). To the mixture was added aqueous KF (10 mL, 1g/10 mL). The mixture was stirred at 25° C. for 30 minutes and extractedwith EtOAc (30 mL×3). The organic layer was dried over Na₂SO₄, filteredand concentrated to yield a residue which was purified by flash silicagel chromatography (from pure PE to PE/EtOAc=3/1, TLC: PE/EtOAc=3/1,R_(f)=0.45) to yield tert-butyl(3R)-3-[[3-acetyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(340 mg, 524.83 μmol, 91.0% yield, 98.9% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.12 (s, 1H), 8.73 (s, 1H), 8.64 (d, J=2.5Hz, 1H), 8.15 (d, J=9.5 Hz, 1H), 8.05 (d, J=8.0 Hz, 1H), 7.28 (s, 1H),7.25-7.19 (m, 1H), 7.18-7.13 (m, 1H), 4.83 (s, 1H), 3.27-3.11 (m, 3H),2.90 (s, 3H), 2.72-2.62 (m, 1H), 2.37 (s, 1H), 2.13-2.06 (m, 1H), 2.05(s, 1H), 1.64 (s, 9H), 1.34 (s, 9H); ES-LCMS m/z 641.4 [M+H]⁺.

Step 4:1-[5-(5-Fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone

To a solution of tert-butyl(3R)-3-[[3-acetyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(340 mg, 524.83 μmol, 1 eq) in DCM (8 mL) was added TFA (3.08 g, 27.01mmol, 2 mL, 51.47 eq). The reaction mixture was stirred at 20° C. for 30min. The reaction mixture was concentrated. The residue was dissolved inwater (10 mL), basified with saturated aqueous NaHCO₃ until pH=8 andextracted with DCM (30 mL×3). The organic layer was dried over Na₂SO₄,filtered and concentrated to yield1-[5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone(250 mg, 515.36 μmol, 98.2% yield, 90.8% purity) as a yellow solid whichwas used in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.11 (s, 1H), 8.58 (d, J=2.9 Hz, 1H), 8.49 (s, 1H), 8.20(td, J=2.4, 9.4 Hz, 1H), 7.90 (s, 1H), 7.44 (d, J=7.9 Hz, 1H), 7.33 (d,J=8.2 Hz, 1H), 7.17 (t, J=7.6 Hz, 1H), 7.13-7.07 (m, 1H), 6.76 (d, J=8.4Hz, 1H), 6.63 (s, 1H), 4.34 (s, 1H), 3.38 (dd, J=4.9, 15.0 Hz, 1H),3.02-2.91 (m, 3H), 2.84 (s, 3H), 2.35 (m, 1H), 2.31-2.23 (m, 1H);ES-LCMS m/z 441.3 [M+H]⁺.

Step 5:(1R)-1-[5-(5-Fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanol (I-198a) &(1S)-1-[5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanol(I-198b)

To a solution of1-[5-(5-fluoro-3-pyridyl)-7-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone(100 mg, 206.14 μmol, 1 eq) in MeOH (2.5 mL) and EtOH (2.5 mL) was addedNaBH₄ (31.19 mg, 824.57 μmol, 4 eq) in portions at 20° C. The reactionmixture was stirred at 20° C. for 0.5 h. H₂O (10 mL) was added to quenchthe reaction and extracted with DCM (30 mL×3). The combined organiclayers were dried over Na₂SO₄, filtered and concentrated to yield aresidue which was purified by preparative HPLC (column: Gemini 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %:55%-70%, 8 min), followed by lyophilization to yield an enantiomer(12.50 mg, 28.25 μmol, 13.7% yield, 100.0% purity) (SFC: Rt=6.742,ee=100%) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.79 (s,1H), 9.34 (s, 1H), 8.66 (d, J=2.8 Hz, 1H), 8.50 (td, J=2.2, 10.4 Hz,1H), 8.12 (s, 1H), 7.99 (d, J=9.3 Hz, 1H), 7.37 (d, J=7.8 Hz, 1H), 7.28(d, J=8.0 Hz, 1H), 7.07 (s, 1H), 7.04-6.98 (m, 1H), 6.97-6.91 (m, 1H),5.20-5.12 (m, 1H), 5.05 (d, J=4.5 Hz, 1H), 4.50 (d, J=8.8 Hz, 1H),3.18-3.00 (m, 2H), 2.98-2.82 (m, 2H), 2.24-2.13 (m, 2H), 1.57 (d, J=6.3Hz, 3H); ES-LCMS m/z 443.2 [M+H]⁺; and the other enantiomer (18.17 mg,40.22 μmol, 19.5% yield, 97.9% purity) (SFC: Rt=5.398, ee=82.9%) as ayellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.75 (s, 1H), 9.30 (s,1H), 8.62 (d, J=2.7 Hz, 1H), 8.50-8.43 (m, 1H), 8.08 (s, 1H), 7.95 (d,J=9.3 Hz, 1H), 7.32 (d, J=7.8 Hz, 1H), 7.24 (d, J=7.8 Hz, 1H), 7.03 (s,1H), 7.00-6.94 (m, 1H), 6.93-6.87 (m, 1H), 5.16-5.08 (m, 1H), 4.99 (d,J=4.4 Hz, 1H), 4.46 (d, J=8.8 Hz, 1H), 3.13-2.98 (m, 2H), 2.92-2.79 (m,2H), 2.14 (m, 2H), 1.53 (d, J=6.4 Hz, 3H); ES-LCMS m/z 443.2 [M+H]⁺.

Example 156

Synthesis of I-200

Synthetic Scheme:

Step 1: 5-Fluoro-N′-(1H-pyrazol-5-yl)pyridine-3-carboxamidine

A mixture of 5-fluoropyridine-3-carbonitrile (1 g, 7.78 mmol, 1 eq) and1H-pyrazol-5-amine (775.80 mg, 9.34 mmol, 1.2 eq) in xylene (15 mL) wasstirred at 70° C. for 30 min, then AlMe₃ (2 M, 7.78 mL, 2 eq) (2 M intoluene, 19.45 mL, 1 eq) was added in one portion at 70° C. The mixturewas stirred at 100° C. for 12 h. The reaction mixture was concentratedunder reduced pressure to yield a residue which was purified by flashsilica gel chromatography (from DCM/MeOH=100/1 to 10/1, TLC:DCM/MeOH=10/1, R_(f)=0.3) to yield5-fluoro-N-(1H-pyrazol-5-yl)pyridine-3-carboxamidine (700 mg, 2.73 mmol,35.1% yield, 80.0% purity) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δppm 8.89 (s, 1H), 8.59 (d, J=2.7 Hz, 1H), 8.06 (d, J=9.3 Hz, 1H), 7.56(d, J=2.2 Hz, 1H), 6.16 (d, J=2.2 Hz, 1H); ES-LCMS m/z 206.3 [M+H]⁺.

Step 2: 2-(5-Fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-ol

A mixture of 5-fluoro-N-(1H-pyrazol-5-yl)pyridine-3-carboxamidine (600mg, 2.34 mmol, 1 eq), CDI (758.62 mg, 4.68 mmol, 2 eq) and DMAP (142.89mg, 1.17 mmol, 0.5 eq) in DMSO (10 mL) was stirred at 130° C. for 12 h.The mixture was purified by preparative HPLC (column: Agela ASB 150*25mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 15%-43%, 9 min),followed by lyophilization to yield2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-ol (150 mg, 627.42μmol, 26.8% yield, 96.7% purity) as a yellow solid. ¹H NMR (400 MHz,CD₃OD) δ ppm 9.17 (s, 1H), 8.82 (d, J=2.4 Hz, 1H), 8.55-8.36 (m, 1H),8.10 (d, J=2.0 Hz, 1H), 6.65 (d, J=2.0 Hz, 1H); ES-LCMS m/z 232.1[M+H]⁺.

Step 3: 4-Chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine

A mixture of 2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-ol(150 mg, 627.42 μmol, 1 eq) and N,N-dimethylaniline (304.12 mg, 2.51mmol, 318.12 μL, 4 eq) in POCl₃ (59.33 g, 386.94 mmol, 35.96 mL, 616.72eq) was degassed and purged with N₂ for 3 times. The mixture was stirredat 110° C. for 3 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to yield a residue which was dilutedwith ice-water, then saturated NaHCO₃ was added to above solution untilpH=8, extracted with EtOAc (30 mL×3). The combined organic layers weredried over Na₂SO₄, filtered and concentrated under reduced pressure toyield a residue which was purified by flash silica gel chromatography(from PE/EtOAC=100/1 to 3/1, TLC: PE/EtOAc=3/1, R_(f)=0.5) to yield4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine (50 mg,172.65 μmol, 27.5% yield, 86.2% purity) as a yellow solid. ¹H NMR (400MHz, CDCl₃) δ ppm 9.51 (s, 1H), 8.65 (d, J=2.7 Hz, 1H), 8.50-8.42 (m,1H), 8.34 (d, J=2.2 Hz, 1H), 6.86 (d, J=2.2 Hz, 1H); ES-LCMS m/z 250.0,252.1 [M+H]⁺.

Step 4:N-[2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-amine (I-200)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazine (50 mg,172.65 μmol, 1 eq), 2-(7-fluoro-2-methyl-1H-indol-3-yl)ethanamine (57.03mg, 224.45 umol, 1.3 eq, HCl) and DIEA (111.57 mg, 863.27 μmol, 150.37μL, 5 eq) in i-PrOH (2 mL) was degassed and purged with N₂ for 3 times.The mixture was stirred at 80° C. for 12 h. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um;mobile phase: [water (0.05% HCl)-ACN]; B %: 55%-85%, 9 min), followed bylyophilization to yieldN-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-amine(19.54 mg, 40.85 μmol, 23.7% yield, 100.0% purity, 2HCl) as a whitesolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.15 (s, 1H), 8.53 (d, J=2.9 Hz,1H), 8.12 (d, J=9.5 Hz, 1H), 8.05 (d, J=2.0 Hz, 1H), 7.33 (d, J=7.8 Hz,1H), 6.86 (dt, J=4.6, 7.8 Hz, 1H), 6.61 (dd, J=7.8, 11.2 Hz, 1H), 6.43(d, J=2.0 Hz, 1H), 3.98 (t, J=6.8 Hz, 2H), 3.13 (t, J=6.8 Hz, 2H), 2.26(s, 3H); ES-LCMS m/z 406.1 [M+H]⁺.

Example 157

Synthesis of I-201

Synthetic Scheme:

Step 1:(3R)—N-2-(5-Fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-201)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazine(11 mg, 24.27 μmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(5.43 mg, 29.13 μmol, 1.2 eq) and DIEA (15.69 mg, 121.37 μmol, 21.14 μL,5 eq) in i-PrOH (2 mL) was stirred at 80° C. for 12 h. The reactionmixture was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Agela ASB 150*25 mm*5 um;mobile phase: [water (0.05% HCl)-ACN]; B %: 60%-90%, 9 min), followed bylyophilization to yield(3R)—N-[2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(4.32 mg, 8.72 μmol, 35.9% yield, 98.2% purity, 2HCl) as a yellow solid.¹H NMR (400 MHz, CD₃OD) δ ppm 9.48 (s, 1H), 8.94 (d, J=9.2 Hz, 1H), 8.82(br s, 1H), 7.96 (s, 1H), 7.34 (d, J=7.6 Hz, 1H), 7.25 (d, J=8.0 Hz,1H), 7.04-6.98 (m, 1H), 6.96-6.90 (m, 1H), 4.86 (m, 1H), 3.25 (d, J=5.6Hz, 1H), 3.12-2.84 (m, 3H), 2.40-2.33 (m, 1H), 2.32 (s, 3H), 2.29-2.17(m, 1H); ES-LCMS m/z 414.2 [M+H]⁺.

Example 158

Synthesis of I-202

Synthetic Scheme:

Step 1: 5-Fluoro-N′-(4-methyl-1H-pyrazol-5-yl)pyridine-3-carboxamidine

A mixture of 5-fluoropyridine-3-carbonitrile (1 g, 6.55 mmol, 1 eq) and4-methyl-1H-pyrazol-5-amine (707.02 mg, 6.55 mmol, 1 eq) in xylene (10mL) was stirred at 70° C. for 0.5 h. Then AlMe₃ (2 M, 3.93 mL, 1.2 eq)was added to the mixture in one portion at 100° C. The mixture wasstirred at 100° C. for 16 h. The mixture was quenched with MeOH (30 mL)and concentrated under reduced pressure. The residue was purified byflash silica gel chromatography (from DCM/MeOH=1/0 to 5/1, TLC:DCM/MeOH=5/1, R_(f)=0.40) to yield product of5-fluoro-N′-(4-methyl-1H-pyrazol-5-yl)pyridine-3-carboxamidine (1.09 g,3.86 mmol, 58.9% yield, 77.7% purity) as yellow oil. ¹H NMR (400 MHz,CD₃OD) δ ppm 8.98 (s, 1H), 8.60 (d, J=2.4 Hz, 1H), 7.38 (s, 1H), 7.17(s, 1H), 2.14 (s, 3H); ES-LCMS m/z 220.2 [M+H]⁺.

Step 2:2-(5-Fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazin-4-ol

A mixture of5-fluoro-N-(4-methyl-1H-pyrazol-5-yl)pyridine-3-carboxamidine (600 mg,2.46 mmol, 1 eq), CDI (799.20 mg, 4.93 mmol, 2 eq) and DMAP (301.07 mg,2.46 mmol, 1 eq) in DMSO (15 mL) was stirred under N₂ atmosphere at 130°C. for 12 h. The reaction mixture was concentrated under reducedpressure. The residue was purified by preparative HPLC (column:Phenomenex Gemini 150*25 mm*10 um; mobile phase: [water(0.05% HCl)-ACN];B %: 14%-44%, 9 min), followed by lyophilization to yield2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazin-4-ol (120mg, 412.82 μmol, 16.8% yield, 96.9% purity, HCl) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 9.12 (s, 1H), 8.72 (s, 1H), 8.34 (s, 1H),7.95 (s, 1H), 2.28 (s, 3H); ES-LCMS m/z 246.1 [M+H]⁺.

Step 3:4-Chloro-2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazine

A mixture of2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazin-4-ol (150mg, 422.79 μmol, 1 eq, 2HCl), N,N-dimethylaniline (69.12 mg, 570.36μmol, 72.30 μL, 1.35 eq) and POCl₃ (11.40 g, 74.38 mmol, 6.91 mL, 175.91eq) was stirred at 110° C. for 3 h. The reaction mixture wasconcentrated under reduced pressure. To the residue was added ice-water(50 mL). The mixture was basified with NaHCO₃ until pH=8 and extractedwith EtOAc (50 mL×3). The organic layer was dried over Na₂SO₄, filteredand concentrated under reduced pressure to yield a residue which waspurified by flash silica gel chromatography (from PE/EtOAc=1/0 to 2/1,TLC: PE/EtOAc=2/1, R_(f)=0.40) to yield4-chloro-2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazine(100 mg, 379.28 μmol, 89.7% yield, 100.0% purity) as a white solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 9.52 (s, 1H), 8.63 (d, J=2.8 Hz, 1H), 8.46(d, J=9.2 Hz, 1H), 8.19 (s, 1H), 2.44 (s, 3H); ES-LCMS m/z 264.0, 266.0[M+H]⁺.

Step 4:N-[2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-202)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazine(20 mg, 75.86 μmol, 1 eq), 2-(7-fluoro-2-methyl-1H-indol-3-yl)ethanamine(21.20 mg, 83.44 μmol, 1.1 eq, HCl) and DIEA (49.02 mg, 379.28 μmol,66.06 μL, 5 eq) in i-PrOH (2 mL) was stirred at 90° C. for 10 h. Themixture was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10um; mobile phase: [water (0.05% HCl)-ACN]; B %: 55%-85%, 9 min) to yieldN-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethyl]-2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazin-4-amine(4.66 mg, 9.21 μmol, 12.1% yield, 97.3% purity, 2HCl) as a yellow solid.¹H NMR (400 MHz, CD₃OD) δ ppm 9.17 (s, 1H), 8.53 (d, J=2.0 Hz, 1H), 8.18(d, J=8.8 Hz, 1H), 7.88 (s, 1H), 7.33 (d, J=7.6 Hz, 1H), 6.92-6.76 (m,1H), 6.60 (dd, J=7.6, 11.2 Hz, 1H), 3.95 (t, J=6.8 Hz, 2H), 3.10 (t,J=6.8 Hz, 2H), 2.27 (s, 3H), 2.23 (s, 3H); ES-LCMS m/z 420.2 [M+H]⁺.

Example 159

Synthesis of I-203

Synthetic Scheme:

Step 1: tert-Butyl N-(4-nitro-1H-pyrazol-5-yl)carbamate

To a solution of 4-nitro-1H-pyrazole-5-carboxylic acid (14 g, 89.12mmol, 1 eq) in toluene (280 mL) was added TEA (22.55 g, 222.81 mmol,31.01 mL, 2.5 eq) and DPPA (26.98 g, 98.04 mmol, 21.24 mL, 1.1 eq) at25° C. The mixture was stirred at 25° C. for 1.5 h. t-BuOH (66.06 g,891.24 mmol, 85.24 mL, 10 eq) was added to the above solution, themixture was stirred at 130° C. for 12 h under N₂. TLC (PE/EtOAc=1/1,R_(f)=0.49) showed the starting material was consumed completely and amajor spot formed. The reaction mixture was quenched by addition ofsaturated NaHCO₃ (100 mL), extracted with EtOAc (80 mL×3). The combinedorganic layers were washed with brine (20 mL), dried over Na₂SO₄,filtered and concentrated under reduced pressure to yield a residuewhich was purified by flash silica gel chromatography (from PE/EtOAc=1/0to 1/1, TLC: PE/EtOAc=1/1, R_(f)=0.49) to yield tert-butylN-(4-nitro-1H-pyrazol-5-yl)carbamate (6.0 g, 21.03 mmol, 23.6% yield,80% purity) as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm13.80-13.47 (m, 1H), 9.35 (br s, 1H), 8.58-7.94 (m, 1H), 1.44 (br s,9H).

Step 2: tert-Butyl N-(4-amino-1H-pyrazol-5-yl)carbamate

A mixture of tert-butyl N-(4-nitro-1H-pyrazol-5-yl)carbamate (5 g, 17.53mmol, 1 eq), Raney-Ni (5 g, in water) and NH₃.H₂O (2.19 g, 17.53 mmol,2.41 mL, 28%, 1 eq) in MeOH (100 mL) was degassed and purged with H₂ for3 times, the mixture was stirred at 20° C. for 2 h under H₂ (15 psi).The reaction mixture was filtered and the filtrate was concentratedunder reduced pressure to yield a crude product of tert-butylN-(4-amino-1H-pyrazol-5-yl)carbamate (3.47 g, 16.44 mmol, 93.7% yield,93.9% purity) as a purple solid which was used in the next step withoutfurther purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.65 (br s, 1H),8.76 (br s, 1H), 7.00 (s, 1H), 3.50 (br s, 2H), 1.50-1.36 (m, 9H);ES-LCMS m/z 143.1 [M-t-Bu+H]⁺.

Step 3: tert-Butyl N-[4-(dimethylamino)-1H-pyrazol-5-yl]carbamate

To a solution of tert-butyl N-(4-amino-1H-pyrazol-5-yl)carbamate (1.5 g,7.11 mmol, 1 eq) in MeOH (30 mL) was added HCHO (938.76 mg, 31.26 mmol,4.4 eq) and NaBH₃CN (4.47 g, 71.06 mmol, 10 eq). The mixture was stirredat 50° C. for 12 h. The reaction mixture was concentrated under reducedpressure to yield a residue which was quenched by addition of water (100mL), extracted with EtOAc (80 mL×3). The combined organic layers werewashed with brine (30 mL), dried over Na₂SO₄, filtered and concentratedunder reduced pressure to yield a residue which was purified by flashsilica gel chromatography (from PE/EtOAc=1/0 to 0/1, TLC: PE/EtOAc=1/1,R_(f)=0.20) to yield tert-butylN-[4-(dimethylamino)-1H-pyrazol-5-yl]carbamate (1.2 g, 3.87 mmol, 54.4%yield, 73.0% purity) as red brown oil. ¹H NMR (400 MHz, CD₃OD) δ ppm7.29 (s, 1H), 2.64 (s, 6H), 1.49 (s, 9H).

Step 4: N4,N4-Dimethyl-1H-pyrazole-4,5-diamine

A mixture of tert-butyl N-[4-(dimethylamino)-1H-pyrazol-5-yl]carbamate(1.2 g, 3.87 mmol, 1 eq) in HCl/MeOH (4 M, 15 mL) was degassed andpurged with N₂ for 3 times, the mixture was stirred at 15° C. for 12 hunder N₂ atmosphere. The reaction mixture was concentrated under reducedpressure to yield a crude product ofN4,N4-dimethyl-1H-pyrazole-4,5-diamine (629 mg, crude, HCl) as a whitesolid which was used in the next step without further purification. ¹HNMR (400 MHz, CD₃OD) δ ppm 8.47 (s, 1H), 3.32 (s, 6H).

Step 5:3-(Dimethylamino)-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol

N4,N4-Dimethyl-1H-pyrazole-4,5-diamine (300 mg, 1.57 mmol, 1 eq, HCl)was dissolved in MeOH (10 mL), then adjusted pH to 9-10 by 2N aq. NaOH.The mixture was concentrated under reduced pressure to yield a residue.The residue and methyl(Z)-3-(5-fluoro-3-pyridyl)-3-hydroxy-prop-2-enoate (332.44 mg, 1.57mmol, 1 eq) in AcOH (15 mL) was stirred at 120° C. for 2 h. The reactionmixture was concentrated under reduced pressure to yield a crude productof 3-(dimethylamino)-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol(616 mg, crude, 2HOAC) as a yellow solid which was used in the next stepwithout further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.71 (s,1H), 9.05 (s, 1H), 8.89 (d, J=1.5 Hz, 1H), 8.30 (s, 1H), 7.97 (s, 1H),6.37 (s, 1H), 2.72-2.52 (m, 6H); ES-LCMS m/z 274.2 [M+H]⁺.

Step 6:7-Chloro-5-(5-fluoro-3-pyridyl)-N,N-dimethyl-pyrazolo[1,5-a]pyrimidin-3-amine

A solution of3-(dimethylamino)-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol(616 mg, 1.57 mmol, 1 eq, 2HOAC) in POCl₃ (16.50 g, 10 mL) was degassedand purged with N₂ for 3 times, the mixture was stirred at 110° C. for 4h under N₂ atmosphere. The reaction mixture was concentrated underreduced pressure to yield a residue which was quenched by addition ofice-water (100 mL) at 0° C., adjusted pH to 9-10 by saturated NaHCO₃,extracted with EtOAc (80 mL×3). The combined organic layers were washedwith brine (20 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to yield a residue which was purified by flash silicagel chromatography (from PE/EtOAc=1/0 to 1/1, TLC: PE/EtOAc=3/1,R_(f)=0.21) to yield7-chloro-5-(5-fluoro-3-pyridyl)-N,N-dimethyl-pyrazolo[1,5-a]pyrimidin-3-amine(190 mg, 651.33 μmol, 41.5% yield, 100% purity) as a red solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.08 (s, 1H), 8.57 (d, J=2.8 Hz, 1H), 8.18-8.10(m, 1H), 7.87 (s, 1H), 7.30 (s, 1H), 3.10 (s, 6H); ES-LCMS m/z 292.2,294.2 [M+H]⁺.

Step 7:5-(5-Fluoro-3-pyridyl)-N7-[2-(1H-indol-3-yl)ethyl]-N3,N3-dimethyl-pyrazolo[1,5-a]pyrimidine-3,7-diamine (I-203)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-N,N-dimethyl-pyrazolo[1,5-a]pyrimidin-3-amine(40 mg, 137.12 μmol, 1 eq) in i-PrOH (5 mL) was added DIEA (53.17 mg,411.37 μmol, 71.65 μL, 3 eq) and 2-(1H-indol-3-yl)ethanamine (28.56 mg,178.26 μmol, 1.3 eq). The mixture was stirred at 90° C. for 12 h. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: PhenomenexGemini C18 250*50 mm*10 μm; mobile phase: [water (0.05% HCl)-ACN]; B %:25%-55%, 9 min), followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-N7-[2-(1H-indol-3-yl)ethyl]-N3,N3-dimethyl-pyrazolo[1,5-a]pyrimidine-3,7-diamine(27.03 mg, 55.35 μmol, 40.3% yield, 100% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.87 (d, J=1.0 Hz, 1H), 8.82 (d,J=1.2 Hz, 1H), 8.36 (s, 1H), 8.17-8.12 (m, 1H), 7.71-7.67 (m, 1H),7.17-7.11 (m, 1H), 7.07-7.00 (m, 2H), 6.98 (s, 1H), 6.18 (s, 1H), 3.96(t, J=6.1 Hz, 2H), 3.48 (s, 6H), 3.21 (t, J=6.0 Hz, 2H); ES-LCMS m/z416.1 [M+H]⁺.

Example 160

Synthesis of I-204a

Synthetic Scheme:

Step 1: (2E)-2-Hydroxyimino-5-methoxy-indan-1-one

A solution of 5-methoxyindan-1-one (5 g, 30.83 mmol, 1 eq) in MeOH (500mL) was heated to 45° C., isopentyl nitrite (5.42 g, 46.24 mmol, 6.23mL, 1.5 eq) and con. HCl (12 M, 5.14 mL, 2 eq) was added. The mixturewas stirred at 45° C. for 2 h. The mixture was concentrate under reducedpressure to yield a residue and MeOH (30 mL) was added. The slurry wasfiltered, dried in vacuo to yield(2E)-2-hydroxyimino-5-methoxy-indan-1-one (5.0 g, 22.75 mmol, 73.8%yield, 87.0% purity) as a brown solid which was used in the next stepwithout further purification. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.47 (s,1H), 7.68 (d, J=8.5 Hz, 1H), 7.14 (d, J=2.0 Hz, 1H), 7.01 (dd, J=2.3,8.5 Hz, 1H), 3.92-3.84 (m, 3H), 3.71 (s, 2H); ES-LCMS m/z 192.1 [M+H]⁺.

Step 2: (2R)-5-Methoxyindan-2-amine

To a solution of (2E)-2-hydroxyimino-5-methoxy-indan-1-one (5 g, 22.75mmol, 1 eq) in AcOH (250 mL) and con. H₂SO₄ (3.5 mL) was added Pd/C (1g, 10%). The mixture was stirred at 25° C. for 12 h under H₂ (15 psi).The mixture was filtered, the filtrate was concentrated under reducedpressure to yield a residue which was poured into ice-water (200 mL),adjusted pH to 10-11 with 2 N NaOH, extracted with EtOAc (150 mL×3). Thecombined organic layers were washed with brine (50 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to yield acrude product of 5-methoxyindan-2-amine (2.0 g, 11.48 mmol, 50.4% yield,93.7% purity) as a brown solid. 5-methoxyindan-2-amine (1 g, 5.74 mmol)was separated by chiral SFC (column: AD-3_EtOH (DEA), AD (250 mm×30 mm,10 μm); mobile phase: [0.1% NH₃H₂O IPA]; B %: 55%-55%, min) for threetimes to yield (2R)-5-methoxyindan-2-amine (220 mg, 1.35 mmol, 23.4%yield, 100% purity) (R_(t)=3.581 min, ee=100%, [α]²² _(D)=24.733 (13.5mg/10 mL, CHCl₃)) as brown oil. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.11 (d,J=8.2 Hz, 1H), 6.78 (s, 1H), 6.72 (dd, J=2.5, 8.3 Hz, 1H), 3.88-3.81 (m,1H), 3.79 (s, 3H), 3.15 (dt, J=6.7, 14.6 Hz, 2H), 2.71-2.59 (m, 2H);ES-LCMS m/z 164.2 [M+H]⁺. And (2S)-5-methoxyindan-2-amine (350 mg, 2.14mmol, 37.3% yield, 100% purity) (R_(t)=3.762 min, ee=100%, [α]²²_(D)=−16.666 (10.3 mg/10 mL, CHCl₃)) as brown oil. ¹H NMR (400 MHz,CDCl₃) δ ppm 7.11 (d, J=7.9 Hz, 1H), 6.78 (s, 1H), 6.72 (dd, J=2.5, 8.3Hz, 1H), 3.87-3.81 (m, 1H), 3.79 (s, 3H), 3.15 (dt, J=6.8, 14.7 Hz, 2H),2.71-2.57 (m, 2H); ES-LCMS m/z 164.2 [M+H]⁺.

Step 3:5-(5-Fluoro-3-pyridyl)-N7-[(2R)-5-methoxyindan-2-yl]-N3,N3-dimethyl-pyrazolo[1,5-a]pyrimidine-3,7-diamine

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-N,N-dimethyl-pyrazolo[1,5-a]pyrimidin-3-amine (50 mg, 171.40 μmol, 1 eq) in i-PrOH (5 mL) was addedDIEA (66.46 mg, 514.21 μmol, 89.56 μL, 3 eq) and(2R)-5-methoxyindan-2-amine (36.37 mg, 222.82 μmol, 1.3 eq). The mixturewas stirred at 70° C. for 12 h. The reaction mixture was concentratedunder reduced pressure to yield a crude5-(5-fluoro-3-pyridyl)-N7-[(2R)-5-methoxyindan-2-yl]-N3,N3-dimethyl-pyrazolo[1,5-a]pyrimidine-3,7-diamine(71 mg, 123.18 μmol, 71.9% yield, 72.6% purity) as red brown oil whichwas used in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.01 (s, 1H), 8.44 (d, J=2.7 Hz, 1H), 8.17-8.10 (m, 1H),7.58 (s, 1H), 7.10 (d, J=8.3 Hz, 1H), 6.75 (s, 1H), 6.71 (dd, J=2.3, 8.2Hz, 1H), 6.43 (d, J=7.6 Hz, 1H), 6.29 (s, 1H), 4.63-4.55 (m, 1H), 3.73(s, 3H), 3.58-3.54 (m, 2H), 3.46-3.37 (m, 2H), 2.93 (s, 6H); ES-LCMS m/z419.2 [M+H]⁺.

Step 4:(2R)-2-[[3-(Dimethylamino)-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol (I-204a)

A solution of5-(5-fluoro-3-pyridyl)-N7-[(2R)-5-methoxyindan-2-yl]-N3,N3-dimethyl-pyrazolo[1,5-a]pyrimidine-3,7-diamine(71 mg, 123.18 μmol, 1 eq) in aq. HBr (5 mL, 60%) was stirred at 120° C.for 2 h under N₂ atmosphere. The reaction mixture was concentrated underreduced pressure to yield a residue which was purified by preparativeHPLC (column: Agela ASB 150*25 mm*5 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 23%-53%, 8 min) followed by lyophilization to yield(2R)-2-[[3-(dimethylamino)-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(33.84 mg, 67.84 μmol, 55.1% yield, 95.7% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.68 (s, 1H), 9.31 (d, J=9.0 Hz,1H), 9.12 (s, 1H), 8.43 (s, 1H), 7.24 (s, 1H), 7.07 (d, J=8.1 Hz, 1H),6.72 (s, 1H), 6.65 (d, J=8.1 Hz, 1H), 4.97-4.94 (m, 1H), 3.54 (s, 6H),3.52-3.43 (m, 2H), 3.19-3.02 (m, 2H); ES-LCMS m/z 405.2 [M+H]⁺.

Example 161

Synthesis of I-204b

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N7-[(2S)-5-methoxyindan-2-yl]-N3,N3-dimethyl-pyrazolo[1,5-a]pyrimidine-3,7-diamine

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-N,N-dimethyl-pyrazolo[1,5-a]pyrimidin-3-amine (50 mg, 171.40 μmol, 1 eq) in i-PrOH (5 mL) was addedDIEA (66.46 mg, 514.20 μmol, 89.56 μL, 3 eq) and(2S)-5-methoxyindan-2-amine (27.98 mg, 171.40 μmol, 1 eq). The mixturewas stirred at 70° C. for 12 h. The reaction mixture was concentratedunder reduced pressure to yield a crude5-(5-fluoro-3-pyridyl)-N7-[(2S)-5-methoxyindan-2-yl]-N3,N3-dimethyl-pyrazolo[1,5-a]pyrimidine-3,7-diamine(71 mg, 128.27 μmol, 74.8% yield, 75.6% purity) as red brown oil whichwas used in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.12-9.05 (m, 1H), 8.52 (d, J=2.7 Hz, 1H), 8.21 (td, J=2.2,9.8 Hz, 1H), 7.66 (s, 1H), 7.18 (d, J=8.3 Hz, 1H), 6.86-6.76 (m, 2H),6.51 (d, J=7.6 Hz, 1H), 6.37 (s, 1H), 4.72-4.57 (m, 1H), 3.81 (s, 3H),3.66 (d, J=4.9 Hz, 2H), 3.50 (ddd, J=7.0, 12.6, 15.9 Hz, 2H), 3.01 (s,6H); ES-LCMS m/z 419.2 [M+H]⁺.

Step 2:(2S)-2-[[3-(Dimethylamino)-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol (I-204b)

A solution of5-(5-fluoro-3-pyridyl)-N7-[(2S)-5-methoxyindan-2-yl]-N3,N3-dimethyl-pyrazolo[1,5-a]pyrimidine-3,7-diamine(71 mg, 128.27 μmol, 1 eq) in aq. HBr (5 mL, 60%) was stirred at 120° C.for 2 h. The reaction mixture was concentrated under reduced pressure toyield a residue which was purified by preparative HPLC (column: AgelaASB 150*25 mm*5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 20%-50%,8 min), followed by lyophilization to yield(2S)-2-[[3-(dimethylamino)-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(32.5 mg, 77.95 μmol, 60.7% yield, 97.0% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 9.64 (s, 1H), 9.25 (d, J=9.3 Hz, 1H), 9.08(s, 1H), 8.43 (s, 1H), 7.22 (s, 1H), 7.08 (d, J=8.1 Hz, 1H), 6.72 (s,1H), 6.65 (d, J=8.1 Hz, 1H), 4.97-4.92 (m, 1H), 3.54 (s, 6H), 3.44-3.41(m, 2H), 3.18-3.03 (m, 2H); ES-LCMS m/z 405.2 [M+H]⁺.

Example 162

Synthesis of I-208

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-208)

A mixture of 7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(60 mg, 225.62 μmol, 1 eq), 2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethanamine(43.65 mg, 270.75 μmol, 1.2 eq), DIEA (87.48 mg, 676.87 μmol, 117.90 μL,3 eq) in i-PrOH (10 mL) was stirred at 80° C. for 12 h. The reactionmixture was concentrated to give the residue which was purified bypreparative HPLC (column: Agela ASB 150*25 mm*5 μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 5%-35%, 9 min), followed by lyophilization toyield5-(5-fluoro-3-pyridyl)-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(37.11 mg, 75.38 μmol, 33.4% yield, 98.7% purity, 3 HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.99 (s, 1H), 8.82 (s, 1H),8.56-8.50 (m, 2H), 8.35 (d, J=8.8 Hz, 1H), 8.26 (s, 1H), 8.10 (s, 1H),7.69-7.63 (m, 1H), 7.04 (s, 1H), 6.65 (d, J=2.0 Hz, 1H), 4.22-4.13 (m,2H), 3.46 (t, J=6.6 Hz, 2H); ES-LCMS m/z 374.2 [M+H]⁺.

Example 163

Synthesis of I-209

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-methyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-209)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (60mg, 228.42 μmol, 1 eq) in i-PrOH (5 mL) was added DIEA (88.56 mg, 685.27μmol, 119.36 μL, 3 eq) and 2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethanamine(45 mg, 279.15 μmol, 1.22 eq). The mixture was stirred at 80° C. for 12h. The reaction mixture was concentrated under reduced pressure to yielda residue which was purified by preparative HPLC (HCl condition; column:Agela ASB 150*25 mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %:15%-45%, 9 min). The desired fraction was lyophilized to yield5-(5-fluoro-3-pyridyl)-3-methyl-N-[2-(1H-pyrrolo[3,2-b]pyridin-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(28.89 mg, 56.64 μmol, 24.8% yield, 97.4% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.94 (s, 1H), 8.79 (d, J=2.7 Hz,1H), 8.57-8.50 (m, 2H), 8.31-8.24 (m, 1H), 8.09 (d, J=7.6 Hz, 2H),7.71-7.63 (m, 1H), 6.80 (s, 1H), 4.12 (t, J=7.0 Hz, 2H), 3.43 (t, J=6.8Hz, 2H), 2.37 (s, 3H); ES-LCMS m/z 388.2 [M+H]⁺.

Example 164

Synthesis of I-210

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-[2-(3-methoxyphenyl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 190.35 μmol, 1 eq) in i-PrOH (3 mL) was added2-(3-methoxyphenyl)ethanamine (43.17 mg, 285.53 μmol, 41.92 μL, 1.5 eq)and DIEA (73.81 mg, 571.06 μmol, 99.47 μL, 3 eq). The mixture wasstirred at 60° C. for 3 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to yield5-(5-fluoro-3-pyridyl)-N-[2-(3-methoxyphenyl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(70 mg, 185.47 μmol, 97.4% yield) as a yellow solid which was used inthe next step without further purification. ¹H NMR (400 MHz, CD₃OD) δppm 9.01 (s, 1H), 8.54 (d, J=2.5 Hz, 1H), 8.25-8.18 (m, 1H), 7.89 (s,1H), 7.18-7.10 (m, 1H), 6.89-6.83 (m, 2H), 6.73-6.65 (m, 1H), 6.36 (s,1H), 3.83 (t, J=6.8 Hz, 2H), 3.67 (s, 3H), 3.03 (t, J=6.8 Hz, 2H), 2.33(s, 3H); ES-LCMS m/z 378.2 [M+H]⁺.

Step 2:3-[2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenol (I-210)

A mixture of5-(5-fluoro-3-pyridyl)-N-[2-(3-methoxyphenyl)ethyl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(70 mg, 185.47 μmol, 1 eq) and aq. HBr (4.47 g, 33.15 mmol, 3 mL, 60%purity, 178.72 eq) was degassed and purged with N₂ for 3 times. Themixture was stirred at 120° C. for 1 h under N₂ atmosphere. The reactionmixture was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Phenomenex Gemini C18 250*50mm*10 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 35%-65%, 10 min),followed by lyophilization to yield3-[2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenol(40.74 mg, 93.37 μmol, 50.3% yield, 100% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CO₃OD) δ ppm 8.87-8.85 (m, 2H), 8.22 (d, J=8.8Hz, 1H), 8.14 (s, 1H), 7.06-6.95 (m, 1H), 6.77-6.68 (m, 2H), 6.60-6.51(m, 1H), 6.44 (s, 1H), 4.01 (t, J=6.5 Hz, 2H), 3.02 (t, J=6.4 Hz, 2H),2.36 (s, 3H); ES-LCMS m/z 364.1 [M+H]⁺.

Example 165

Synthesis of I-211

Synthetic Scheme:

Step 1:N-[2-(4-Aminophenyl)ethyl]-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-211)

To a solution of 4-(2-aminoethyl)aniline (25.92 mg, 190.35 μmol, 1 eq)in i-PrOH (3 mL) was added7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 190.35 μmol, 1 eq) and DIEA (73.81 mg, 571.06 μmol, 99.47 μL, 3 eq).The mixture was stirred at 50° C. for 12 h. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Agela ASB 150*25 mm*5 μm; mobilephase: [water (0.05% HCl)-ACN]; B %: 20%-50%, 8 min) followed bylyophilization to yieldN-[2-(4-aminophenyl)ethyl]-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine (73.99 mg, 155.10 μmol, 81.5% yield, 98.9% purity,3HCl) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.94 (s, 1H),8.80 (d, J=2.7 Hz, 1H), 8.28 (td, J=2.4, 8.9 Hz, 1H), 8.11 (s, 1H), 7.54(d, J=8.6 Hz, 2H), 7.35 (d, J=8.3 Hz, 2H), 6.80 (s, 1H), 4.02 (t, J=7.2Hz, 2H), 3.19 (t, J=7.2 Hz, 2H), 2.37 (s, 3H); ES-LCMS m/z 363.3 [M+H]⁺.

Example 166

Synthesis of I-213

Synthetic Scheme:

Step1:4-[2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenol (I-213)

To a mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (60mg, 228.42 μmol, 1 eq) and 4-(2-aminoethyl)phenol (37.60 mg, 274.11μmol, 1.2 eq) in i-PrOH (12 mL) was added DIEA (88.57 mg, 685.27 μmol,119.36 μL, 3 eq). The mixture was stirred at 75° C. for 19 h under N₂atmosphere. The reaction mixture was concentrated under reduced pressureto yield a residue which was purified by preparative HPLC (column: AgelaASB 150*25 mm*5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 30%-60%,9 min), followed by lyophilization to yield4-[2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]phenol(55.37 mg, 126.91 μmol, 55.6% yield, 100.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.84 (br s, 2H), 8.23-8.12 (m, 2H),7.06 (d, J=8.6 Hz, 2H), 6.63-6.57 (m, 2H), 6.36 (s, 1H), 3.96 (t, J=6.5Hz, 2H), 2.99 (t, J=6.5 Hz, 2H), 2.36 (s, 3H); ES-LCMS m/z 364.0 [M+H]⁺.

Example 167

Synthesis of I-214

Synthetic Scheme:

Step 1:N-[2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-amine(I-214)

To a mixture of 7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine(100 mg, 376.04 μmol, 1 eq) in DIEA (5 mL) and i-PrOH (3 mL) was added2-(7-fluoro-2-methyl-1H-indol-3-yl)ethanamine (95.55 mg, 376.04 μmol, 1eq, HCl). The mixture was stirred at 80° C. for 12 h. The reactionmixture was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Gemini 150*25 5 u; mobilephase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 45%-75%, 8 min),followed by lyophilization to yieldN-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-amine(58.73 mg, 141.11 μmol, 37.5% yield, 97.2% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.70 (s, 1H), 8.44 (d, J=2.8 Hz, 1H), 7.93(d, J=2.4 Hz, 1H), 7.81-7.76 (m, 1H), 7.25 (d, J=7.6 Hz, 1H), 6.99 (dt,J=4.8, 8.0 Hz, 1H), 6.83-6.79 (m, 1H), 6.47 (d, J=2.4 Hz, 1H), 5.99 (s,1H), 3.72 (q, J=6.4 Hz, 2H), 3.11 (t, J=6.4 Hz, 2H), 2.24 (s, 3H);ES-LCMS m/z 405.2 [M+H]⁺.

Example 168

Synthesis of I-215

Synthetic Scheme:

Step 1:3-[2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-1H-indol-5-ol(I-215)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (60mg, 228.42 μmol, 1 eq), 3-(2-aminoethyl)-1H-indol-5-ol (60.37 mg, 283.87μmol, 1.24 eq, HCl) in i-PrOH (5 mL) was added DIEA (147.61 mg, 1.14μmol, 198.94 μL, 5 eq). The mixture was stirred at 60° C. for 4 h. Thereaction mixture was concentrated to yield a residue which was purifiedon silica gel column chromatography (from PE/EtOAc=1/0 to 1/1, TLC:PE/EtOAc=1/1, R_(f)=0.25) to yield the product3-[2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]-1H-indol-5-ol(73.19 mg, 179.69 μmol, 78.7% yield, 98.8% purity) as a yellow solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 10.52 (br s, 1H), 9.16 (s, 1H), 8.77-8.55(m, 2H), 8.27 (d, J=9.7 Hz, 1H), 8.07-7.93 (m, 2H), 7.20-7.02 (m, 2H),6.91 (d, J=1.8 Hz, 1H), 6.65 (s, 1H), 6.62-6.58 (m, 1H), 3.91-3.74 (m,2H), 3.03 (t, J=7.1 Hz, 2H), 2.28 (s, 3H); ES-LCMS m/z 403.2 [M+H]⁺.

Example 169

Synthesis of I-216a

Synthetic Scheme:

Step 1:(3R)-8-Fluoro-N-[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-216a)

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 190.35 μmol, 1 eq),(3R)-8-fluoro-2,3,4,9-tetrahydro-1H-carbazol-3-amine (47.36 mg, 209.39μmol, 1.1 eq) and DIEA (123.01 mg, 951.76 μmol, 165.78 μL, 5 eq) ini-PrOH (2 mL) was stirred at 90° C. for 12 h. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Phenomenex Gemini C18 250*50 mm*10um; mobile phase: [water (0.05% HCl)-ACN]; B %: 50%-80%, 10 min). Thedesired fraction was lyophilized to yield(3R)-8-fluoro-N-[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(21.98 mg, 51.06 μmol, 26.8% yield, 100.0% purity) as a yellow solid(SFC: Rt=2.106, ee %=97.3%). ¹H NMR (400 MHz, CD₃OD) δ ppm 8.97 (s, 1H),8.75 (d, J=2.4 Hz, 1H), 8.29 (td, J=2.0, 9.2 Hz, 1H), 8.13 (s, 1H), 7.16(d, J=8.0 Hz, 1H), 7.00 (s, 1H), 6.89 (dt, J=4.8, 8.0 Hz, 1H), 6.76 (dd,J=8.0, 11.6 Hz, 1H), 4.62 (m, 1H), 3.32 (s, 1H), 3.16-3.06 (m, 1H),3.03-2.95 (m, 2H), 2.37 (s, 3H), 2.36-2.23 (m, 2H); ES-LCMS m/z 431.2[M+H]⁺.

Example 170

Synthesis of I-218

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-methyl-N-(2-phenylethy)pyrazolo[1,5-a]pyrimidin-7-amine(I-218)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 190.35 μmol, 1 eq) in i-PrOH (3 mL) was added DIEA (73.80 mg, 571.06μmol, 99.47 μL, 3 eq) and 2-phenylethanamine (34.60 mg, 285.53 μmol,35.86 μL, 1.5 eq). The mixture was stirred at 60° C. for 12 h. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: Agela ASB 150*25mm*5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 45%-75%, 9 min),followed by lyophilization to yield5-(5-fluoro-3-pyridyl)-3-methyl-N-(2-phenylethyl)pyrazolo[1,5-a]pyrimidin-7-amine(37.43 mg, 89.05 μmol, 46.78% yield, 100% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.83-8.78 (m, 2H), 8.17-8.13 (m,2H), 7.32-7.27 (m, 2H), 7.23 (t, J=7.3 Hz, 2H), 7.18-7.13 (m, 1H), 6.51(s, 1H), 4.01 (t, J=6.7 Hz, 2H), 3.09 (t, J=6.7 Hz, 2H), 2.34 (s, 3H);ES-LCMS m/z 348.2 [M+H]⁺.

Example 171

Synthesis of I-219

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-[(2S)-5-methoxyindan-2-yl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 190.35 μmol, 1 eq), (2S)-5-methoxyindan-2-amine (33.05 mg, 190.35μmol, 1 eq), DIEA (73.80 mg, 571.06 μmol, 99.47 μL, 3 eq) in i-PrOH (10mL) was stirred at 80° C. for 12 h. The reaction mixture wasconcentrated to yield5-(5-fluoro-3-pyridyl)-N-[(2S)-5-methoxyindan-2-yl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(60 mg, 122.03 μmol, 64.1% yield, 79.2% purity) as brown oil which wasused in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.06-8.95 (m, 1H), 8.58-8.43 (m, 1H), 8.22-8.09 (m, 1H),7.79 (s, 1H), 7.15-7.00 (m, 1H), 6.80-6.61 (m, 2H), 6.53 (d, J=7.1 Hz,1H), 6.32 (s, 1H), 4.59 (s, 1H), 3.74 (s, 3H), 3.60 (d, J=6.4 Hz, 2H),3.06-3.02 (m, 2H), 2.31 (s, 3H); ES-LCMS m/z 390.2 [M+H]⁺.

Step 2:(2S)-2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol (I-219)

5-(5-Fluoro-3-pyridyl)-N-[(2S)-5-methoxyindan-2-yl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(60 mg, 122.03 μmol, 1 eq) in HBr (5 mL, 60% in water) was stirred at120° C. for 2 h. The reaction mixture was concentrated to yield aresidue which was purified by preparative HPLC (column: PhenomenexGemini C18 250*50 mm*10 μm; mobile phase: [water (0.05% HCl)-ACN]; B %:30%-60%, 10 min), followed by lyophilization to yield(2S)-2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(23.51 mg, 52.44 μmol, 43.0% yield, 100.0% purity, 2 HCl) ((SFC:Rt=2.090, ee=100%) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.08(s, 1H), 8.90 (s, 1H), 8.45 (d, J=9.0 Hz, 1H), 8.16 (s, 1H), 7.09 (d,J=8.0 Hz, 1H), 7.01 (s, 1H), 6.73 (s, 1H), 6.66 (dd, J=2.0, 8.0 Hz, 1H),5.11-5.02 (m, 1H), 3.54-3.42 (m, 2H), 3.28-3.14 (m, 2H), 2.41 (s, 3H);ES-LCMS m/z 376.2 [M+H]⁺.

Example 172

Synthesis of I-220

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[(2S)-5-methoxyindan-2-yl]pyrazolo[1,5-a]pyrimidin-7-amine

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(50 mg, 154.79 μmol, 1 eq), (2S)-5-methoxyindan-2-amine (26.88 mg,154.79 μmol, 1 eq) and DIEA (60.02 mg, 464.36 μmol, 80.88 μL, 3 eq) ini-PrOH (10 mL) was stirred at 80° C. for 12 h. The reaction mixture wasconcentrated to yield5-(5-fluoro-3-pyridyl)-3-isopropyl-N-[(2S)-5-methoxyindan-2-yl]pyrazolo[1,5-a]pyrimidin-7-amine(60 mg, 115.69 μmol, 74.7% yield, 80.5% purity) as brown oil which wasused in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.06-8.93 (m, 1H), 8.54-8.42 (m, 1H), 8.13 (d, J=7.3 Hz,1H), 7.79 (s, 1H), 7.10 (d, J=7.9 Hz, 1H), 6.79-6.58 (m, 2H), 6.51 (d,J=7.3 Hz, 1H), 6.33 (s, 1H), 4.57 (s, 1H), 3.75-3.62 (m, 3H), 3.59 (d,J=6.8 Hz, 2H), 3.02 (d, J=8.6 Hz, 3H), 1.15 (d, J=2.2 Hz, 6H); ES-LCMSm/z 418.2[M+H]⁺.

Step 2:(2S)-2-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol (I-220)

5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[(2S)-5-methoxyindan-2-yl]pyrazolo[1,5-a]pyrimidin-7-amine(60 mg, 115.69 μmol, 1 eq) in HBr (5 mL, 60% in water) was stirred at120° C. for 2 h. The reaction mixture was concentrated to yield theresidue which was purified by preparative HPLC (column: PhenomenexGemini C18 250*50 mm*10 um; mobile phase: [water (0.05% HCl)-ACN]; B %:40%-70%, 10 min) followed by lyophilization to yield(2S)-2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(17.20 mg, 36.11 μmol, 31.2% yield, 100.0% purity, 2 HCl) (SFC:Rt=2.050, ee=99.374%) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm9.04 (s, 1H), 8.86 (d, J=2.0 Hz, 1H), 8.40 (d, J=9.0 Hz, 1H), 8.26 (s,1H), 7.09 (d, J=8.0 Hz, 1H), 6.99 (s, 1H), 6.73 (s, 1H), 6.66 (dd,J=2.5, 8.0 Hz, 1H), 5.09-5.01 (m, 1H), 3.54-3.43 (m, 2H), 3.39 (td,J=6.8, 13.9 Hz, 1H), 3.26-3.15 (m, 2H), 1.41 (d, J=7.0 Hz, 6H); ES-LCMSm/z 404.2 [M+H]⁺.

Example 173

Synthesis of I-222a, I-222b and I-222c

Step 1: 7-Chloro-5-(5-fluoro-3-pyridyl)-3-iodo-pyrazolo[1,5-a]pyrimidine

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine (600 mg, 2.24mmol, 1 eq) in DCM (25 mL) and MeCN (25 mL) was added NIS (1.01 g, 4.48mmol, 2 eq) partwise and the mixture was stirred at 20° C. for 24 h. TLC(PE/EtOAc=3/1, R_(f)=0.41) showed the starting material was consumedcompletely. The reaction mixture was quenched with saturated Na₂S203(100 mL) and concentrated under reduced pressure to remove DCM and MeCN.The residue was extracted with EtOAc (100 mL×3). The organic layer wasdried over Na₂SO₄, filtered and concentrated under reduced pressure toyield a residue which was purified by flash silica gel chromatography(from PE/EtOAc=1/0 to 4/1, TLC: PE/EtOAc=3/1, R_(f)=0.41) to yield7-chloro-5-(5-fluoro-3-pyridyl)-3-iodo-pyrazolo[1,5-a]pyrimidine (1.03g, 2.75 mmol, 61.4% yield, 100.0% purity) as a yellow solid. ¹H NMR (400MHz, CDCl₃) δ ppm 9.12 (s, 1H), 8.64 (d, J=2.8 Hz, 1H), 8.34-8.26 (m,2H), 7.53-7.45 (m, 1H); ES-LCMS m/z 374.9 [M+H]⁺.

Step 2:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-iodo-pyrazolo[1,5-a]pyrimidin-7-amine

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-iodo-pyrazolo[1,5-a]pyrimidine (650mg, 1.74 mmol, 1 eq), 2-(1H-indol-3-yl)ethanamine (333.66 mg, 2.08 mmol,1.2 eq) and DIEA (672.89 mg, 5.21 mmol, 906.86 μL, 3 eq) in i-PrOH (50mL) was stirred at 60° C. for 12 h. TLC (PE/EtOAc=2/1, R_(f)=0.30)showed the starting material was consumed completely. The reactionmixture was cooled to 20° C. and filtered. The solid was washed withi-PrOH (5 mL), dried under reduced pressure to yield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-iodo-pyrazolo[1,5-a]pyrimidin-7-amine(620 mg, 1.24 mmol, 71.3% yield, 99.3% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.78 (s, 1H), 8.52 (d, J=2.4 Hz, 1H), 8.10-7.90(m, 3H), 7.68 (d, J=7.6 Hz, 1H), 7.39-7.32 (m, 1H), 7.24-7.18 (m, 2H),7.06 (s, 1H), 6.57 (br s, 1H), 6.12 (s, 1H), 3.86 (q, J=6.4 Hz, 2H),3.27 (t, J=6.4 Hz, 2H); ES-LCMS m/z 499.1 [M+H]⁺.

Step 3: tert-Butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)-3-iodo-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate

A mixture of5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-iodo-pyrazolo[1,5-a]pyrimidin-7-amine(900 mg, 1.81 mmol, 1 eq), Boc₂O (1.58 g, 7.22 mmol, 1.66 mL, 4 eq) andDMAP (1.10 g, 9.03 mmol, 5 eq) in 1,4-dioxane (60 mL) was stirred at120° C. for 12 h. TLC (PE/EtOAc=3/1, R_(f)=0.60) showed the startingmaterial was consumed completely. The reaction mixture was concentratedunder reduced pressure to yield a residue which was purified by flashsilica gel chromatography (from PE/EtOAc=1/0 to 5/1, TLC: PE/EtOAc=3/1,R_(f)=0.60) to yield tert-butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)-3-iodo-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(1 g, 1.41 mmol, 78.2% yield, 98.7% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.73 (s, 1H), 8.55 (d, J=2.4 Hz, 1H), 8.15 (s,1H), 7.98 (br s, 1H), 7.90 (d, J=7.6 Hz, 1H), 7.42 (d, J=7.6 Hz, 1H),7.25-7.23 (m, 2H), 7.19-7.12 (m, 1H), 6.55 (s, 1H), 4.24-4.18 (m, 2H),3.15-3.06 (m, 2H), 1.59 (s, 9H), 1.39 (s, 9H); ES-LCMS m/z 699.2 [M+H]⁺.

Step 4: tert-Butyl3-[2-[[3-acetyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate

A mixture of tert-butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)-3-iodo-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(200 mg, 282.60 μmol, 1 eq), tributyl(1-ethoxyvinyl)stannane (880 mg,2.44 mmol, 822.43 μL, 8.62 eq) and Pd(dppf)Cl₂ (206.78 mg, 282.60 μmol,1 eq) in toluene (6 mL) was bubbled with N₂ for 2 minutes and sealed.The reaction mixture was irradiated under microwave (1 bar) at 100° C.for 2 h. The reactions were carried out in parallel 5 times. KF (100 mL,2M) was added to the combined mixture. The mixture was stirred at 25° C.for 1 h and extracted with EtOAc (100 mL×3). The organic layer was driedover Na₂SO₄, filtered and concentrated under reduced pressure to yield aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 4/1, TLC: PE/EtOAc=3/1, R_(f)=0.40) to yield tert-butyl3-[2-[[3-acetyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate(450 mg, 732.11 μmol, 51.8% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.79 (s, 1H), 8.66 (s, 1H), 8.61 (d, J=2.4Hz, 1H), 7.97 (d, J=6.4 Hz, 1H), 7.80 (td, J=2.4, 9.2 Hz, 1H), 7.42 (d,J=7.6 Hz, 1H), 7.26-7.20 (m, 2H), 7.17-7.11 (m, 1H), 6.65 (s, 1H), 4.30(t, J=6.4 Hz, 2H), 3.18-3.12 (m, 2H), 2.84 (s, 3H), 1.59 (s, 9H), 1.43(s, 9H); ES-LCMS m/z 615.3 [M+H]⁺.

Step 5:1-[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone

To a mixture of tert-butyl3-[2-[[3-acetyl-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate(450 mg, 732.11 μmol, 1 eq) in MeOH (50 mL) was added HCl/H₂O (6 M, 150mL, 1229.33 eq) at 20° C. The mixture was stirred at 35° C. for 3 h. Themixture was basified with aqueous NaOH (15%) until pH=8 and concentratedunder reduced pressure to remove MeOH. The residue was extracted withEtOAc (50 mL×3). The organic layer was dried over Na₂SO₄, filtered andconcentrated under reduced pressure to yield1-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone(300 mg, 723.88 μmol, 98.9% yield, 100.0% purity) as a yellow solidwhich was used in the next step without further purification. ¹H NMR(400 MHz, DMSO-d₆) δ ppm 10.77 (br s, 1H), 9.11 (s, 1H), 8.67 (d, J=2.4Hz, 1H), 8.60 (t, J=6.0 Hz, 1H), 8.49 (s, 1H), 8.18 (d, J=10.4 Hz, 1H),7.61 (d, J=7.6 Hz, 1H), 7.24 (d, J=7.6 Hz, 1H), 7.16 (d, J=1.6 Hz, 1H),7.05-6.99 (m, 1H), 6.98-6.93 (m, 1H), 6.82 (s, 1H), 3.84 (q, J=6.4 Hz,2H), 3.09 (t, J=6.8 Hz, 2H), 2.67 (s, 3H); ES-LCMS m/z 415.1 [M+H]⁺.

Step 6:(1S)-1-[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanol(I-222b) &(1R)-1-[5-(5-Fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanol(I-222a)

To a solution of1-[5-(5-fluoro-3-pyridyl)-7-[2-(1H-indol-3-yl)ethylamino]pyrazolo[1,5-a]pyrimidin-3-yl]ethanone(250 mg, 603.23 μmol, 1 eq) in THF (50 mL) and EtOH (20 mL) was addedNaBH₄ (114.11 mg, 3.02 mmol, 5 eq) partwise at 20° C. The mixture wasstirred at 20° C. for 3 h. The reaction mixture was quenched with water(50 mL) and concentrated under reduced pressure at 30° C. to remove EtOHand THF. The residue was extracted with EtOAc (80 mL×3), dried overNa₂SO₄, filtered and concentrated under reduced pressure to yield aresidue which was separated by chiral SFC (column: DAICEL CHIRALPAK AD-H(250 mm*30 mm, 5 um); mobile phase: [0.1% NH₃.H₂O/EtOH]; B %: 40%-40%).Peak 1 was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10um; mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %:40%-70%, 10 min). The desired fraction was lyophilized to yield anenantiomer (32.78 mg, 76.81 μmol, 12.7% yield, 97.6% purity) as a whitesolid (SFC: Rt=6.825, ee=98.8%; Optical Rotation: [a]^(22.3) _(D)=−9.540(10.04 mg/10 mL in MeOH)). ¹H NMR (400 MHz, CDCl₃) δ ppm 8.78 (s, 1H),8.51 (d, J=2.4 Hz, 1H), 8.07 (br s, 1H), 7.95 (s, 1H), 7.87-7.80 (m,1H), 7.68 (d, J=7.6 Hz, 1H), 7.40-7.32 (m, 1H), 7.26-7.18 (m, 2H), 7.06(s, 1H), 6.56 (br s, 1H), 6.08 (s, 1H), 5.34 (d, J=6.0 Hz, 1H),3.89-3.83 (m, 2H), 3.29-3.24 (m, 3H), 1.70 (d, J=6.4 Hz, 3H); ES-LCMSm/z 417.2 [M+H]⁺. Peak 2 was concentrated under reduced pressure toyield a residue which was purified by preparative HPLC (column:Phenomenex Gemini 150*25 mm*10 um; mobile phase: [water (0.05% ammoniahydroxide v/v)-ACN]; B %: 40%-70%, 10 min). The desired fraction waslyophilized to yield the residue which was separated by chiral SFC(column: DAICEL CHIRALPAK AD-H (250 mm*30 mm, 5 um); mobile phase: [0.1%NH₃.H₂O/EtOH]; B %: 40%-40%) again. The desired fraction wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 42%-72%,10 min). The desired fraction was lyophilized to yield the otherenantiomer (30.13 mg, 72.35 μmol, 12.0% yield, 100.0% purity) as a whitesolid (SFC: Rt=7.042, ee=99.0%; Optical Rotation: [a]^(22.2) _(D)=1.916(6.44 mg/10 mL in MeOH)). ¹H NMR (400 MHz, CDCl₃) δ ppm 8.77 (s, 1H),8.51 (d, J=2.4 Hz, 1H), 8.11 (br s, 1H), 7.95 (s, 1H), 7.83 (td, J=2.0,9.6 Hz, 1H), 7.68 (d, J=7.6 Hz, 1H), 7.36 (d, J=8.0 Hz, 1H), 7.27-7.18(m, 2H), 7.06 (d, J=2.0 Hz, 1H), 6.58 (t, J=5.6 Hz, 1H), 6.07 (s, 1H),5.42-5.26 (m, 1H), 3.85 (q, J=6.4 Hz, 2H), 3.34-3.22 (m, 3H), 1.70 (d,J=6.4 Hz, 3H); ES-LCMS m/z 417.2 [M+H]⁺.

Example 174

Synthesis of I-224

Synthetic Scheme:

Step 1:N-[2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo(I-224)

A mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (60mg, 228.42 μmol, 1 eq) and 2-(7-fluoro-2-methyl-1H-indol-3-yl)ethanamine(79.89 mg, 296.95 μmol, 1.3 eq, HCl) in i-PrOH (10 mL) was added DIEA(88.57 mg, 685.27 μmol, 119.36 μL, 3 eq) and purged with N₂ for 3 times.The mixture was stirred at 80° C. for 16 h under N₂ atmosphere. Thereaction mixture was concentrated under reduced pressure to give aresidue which was purified by preparative HPLC (column: Agela DurashellC18 150*25 5 u; mobile phase: [water (0.05% HCl)-ACN]; B %: 30%-60%, 12min) to yield a product which was purified by preparative HPLC (column:Gemini 150*25 5 u; mobile phase: [water (0.05% ammonia hydroxidev/v)-ACN]; B %: 55%-85%, 8 min), followed by lyophilization to yieldN-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(16.79 mg, 40.13 μmol, 17.6% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.80 (s, 1H), 8.50 (d, J=2.6 Hz, 1H), 8.00(br s, 1H), 7.95-7.82 (m, 2H), 7.32 (d, J=7.9 Hz, 1H), 7.10-7.03 (m,1H), 6.89 (dd, J=8.2, 11.0 Hz, 1H), 6.49 (br s, 1H), 6.01 (s, 1H), 3.78(q, J=6.5 Hz, 2H), 3.21-3.13 (m, 2H), 2.38 (s, 3H), 2.29 (s, 3H);ES-LCMS m/z 419.2 [M+H]⁺.

Example 175

Synthesis of I-225

Synthetic Scheme:

Step 1: Methyl7-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethylamino]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

To a solution of 2-(7-fluoro-2-methyl-1H-indol-3-yl)ethanamine (50 mg,196.77 μmol, 1 eq, HCl) in/i-PrOH (5 mL) was added methyl7-chloro-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(63.52 mg, 196.77 μmol, 1 eq) and DIEA (76.29 mg, 590.31 μmol, 102.82μL, 3 eq). The mixture was stirred at 60° C. for 3 h under N₂atmosphere. The mixture was concentrated under reduced pressure andwater (50 mL) was added. The mixture was extracted with EtOAc (30 mL×3).The combined organic layers were washed with brine (20 mL), dried overNa₂SO₄, filtered and concentrated to yield methyl7-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethylamino]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(90 mg, 173.79 μmol, 88.3% yield, 89.3% purity) as a yellow solid whichwas used in the next step without further purification. ¹H NMR (400 MHz,CD₃OD) δ ppm 8.51-8.46 (m, 2H), 8.41 (s, 1H), 7.72-7.65 (m, 1H), 7.38(d, J=8.0 Hz, 1H), 6.98-6.92 (m, 1H), 6.69-6.60 (m, 1H), 5.85 (s, 1H),3.88 (s, 3H), 3.83 (br s, 2H), 3.16-3.07 (m, 2H), 2.08 (s, 3H); ES-LCMSm/z 463.2 [M+H]⁺.

Step 2:7-[2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethylamino]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid

To a solution of methyl7-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethylamino]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylate(90 mg, 173.79 μmol, 1 eq) in MeOH (5 mL) was added NaOH (2 M, 4.96 mL,57.09 eq). The mixture was stirred at 50° C. for 4 h under N₂atmosphere. The mixture was concentrated under reduced pressure. Theresidue was dissolved in water (50 mL), adjusted to pH to 3 by 1N HClsolution, extracted with EtOAc (20 mL×3). The combined organic layerswere washed with brine (20 mL), dried over Na₂SO₄, filtered andconcentrated to yield7-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethylamino]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (80 mg, 160.56 μmol, 92.4% yield, 90.0% purity) as a yellow solidwhich was used in the next step without further purification. ¹H NMR(400 MHz, CD₃OD) δ ppm 8.48 (s, 2H), 8.41 (s, 1H), 7.68 (d, J=9.8 Hz,1H), 7.38 (d, J=7.8 Hz, 1H), 6.95 (dt, J=4.5, 7.9 Hz, 1H), 6.68-6.60 (m,1H), 5.85 (s, 1H), 3.89-3.83 (m, 2H), 3.15-3.08 (m, 2H), 2.08 (s, 3H);ES-LCMS m/z 449.2 [M+H]⁺.

Step 3:[7-[2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethylamino]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-3-yl]-morpholino-methanone(I-225)

To a solution of7-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethylamino]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidine-3-carboxylicacid (80 mg, 160.56 μmol, 1 eq) in DCM (10 mL) was added morpholine(20.98 mg, 240.84 μmol, 21.19 μL, 1.5 eq), HATU (91.58 mg, 240.84 μmol,1.5 eq) and TEA (32.49 mg, 321.12 μmol, 44.70 μL, 2 eq). The mixture wasstirred at 20° C. for 1 h under N₂ atmosphere. The mixture wasconcentrated under reduced pressure, water (30 mL) was added. Themixture was extracted with EtOAc (20 mL×3). The combined organic layerswere washed with brine (30 mL), dried over Na₂SO₄, filtered andconcentrated to yield a residue which was purified by preparative HPLC(column: Phenomenex Gemini C18 250*50 mm*10 μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 33%-63%, 10 min), followed by lyophilization toyield[7-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethylamino]-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-3-yl]-morpholino-methanone(29.09 mg, 56.21 μmol, 35.0% yield, 100% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 8.78 (d, J=2.5 Hz, 1H), 8.60 (s, 1H), 8.38(s, 1H), 7.79-7.70 (m, 1H), 7.34 (d, J=8.0 Hz, 1H), 6.87 (dt, J=4.7, 7.8Hz, 1H), 6.60 (dd, J=7.7, 11.4 Hz, 1H), 5.88 (s, 1H), 4.01-3.93 (m, 2H),3.76 (s, 8H), 3.20-3.14 (m, 2H), 2.12 (s, 3H); ES-LCMS m/z 518.3 [M+H]⁺.

Example 176

Synthesis of I-226

Synthetic Scheme:

Step 1: tert-Butyl3-[2-[[3-bromo-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate

To a solution of3-bromo-5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(750 mg, 1.63 mmol, 1 eq) in 1,4-dioxane (30 mL) was added DMAP (596.31mg, 4.88 mmol, 3 eq) and (Boc)₂O (887.73 mg, 4.07 mmol, 934.45 μL, 2.5eq). The mixture was stirred at 110° C. for 5 h. The reaction mixturewas concentrated under reduced pressure to yield a residue which waspurified by flash silica gel chromatography (from PE/EtOAc=100/1 to 3/1,TLC: PE/EtOAc=3/1, R_(f)=0.75) to yield tert-butyl3-[2-[[3-bromo-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate(950 mg, 1.46 mmol, 89.6% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl3) δ ppm 8.73 (s, 1H), 8.57 (d, J=2.8 Hz, 1H), 8.16(s, 1H), 8.00 (d, J=7.0 Hz, 1H), 7.91 (td, J=2.1, 9.3 Hz, 1H), 7.45 (d,J=7.8 Hz, 1H), 7.27-7.21 (m, 2H), 7.21-7.14 (m, 1H), 6.57 (s, 1H), 4.24(t, J=6.8 Hz, 2H), 3.12 (t, J=6.7 Hz, 2H), 1.62 (s, 9H), 1.42 (s, 9H);ES-LCMS m/z 651.2, 652.2 [M+H]⁺.

Step 2: tert-Butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)-3-tetrahydrofuran-2-yl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate

4-tert-Butyl-2-(4-tert-butyl-2-pyridyl)pyridine (24.72 mg, 92.09 μmol,0.1 eq) and dichloronickel; 1,2-dimethoxyethane (20.23 mg, 92.09 μmol,0.1 eq) were added into THE (4 mL). The mixture was stirred at 50° C.under N₂ atmosphere until a green solution was obtained.[Ir{dFCF₃ppy}₂(bpy)]PF₆ (51.66 mg, 46.05 μmol, 0.05 eq), K₂HPO₄ (320.81mg, 1.84 mmol, 2 eq) and a solution of tert-butyl3-[2-[[3-bromo-5-(5-fluoro-3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate(600 mg, 920.92 μmol, 1 eq) in TH (20 mL) were added into the abovemixture under N₂ atmosphere. The resulting mixture was stirred andirradiated with a standard 72 W LED strip light bulb at 25° C. for 12 h.The reaction mixture was concentrated to yield a residue which waspurified by flash silica gel chromatography (from PE/EtOAc=100/1 to 3/1,TLC: PE/EtOAc=3/1, R_(f)=0.45) to yield tert-butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)-3-tetrahydrofuran-2-yl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(190 mg, 274.04 μmol, 29.8% yield, 92.7% purity) as yellow oil. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.83-8.76 (m, 1H), 8.55 (s, 1H), 8.18 (s, 1H),8.05 (s, 1H), 7.90-7.79 (m, 1H), 7.47 (t, J=6.9 Hz, 1H), 7.31 (d, J=8.8Hz, 1H), 7.19 (t, J=7.6 Hz, 2H), 6.59 (s, 1H), 5.38-5.29 (m, 1H),4.21-4.15 (m, 2H), 4.10 (m, 1H), 4.01-3.92 (m, 1H), 3.11 (t, J=6.9 Hz,2H), 2.45-2.35 (m, 1H), 2.33-2.24 (m, 1H), 2.23-2.16 (m, 1H), 2.15-2.07(m, 1H), 1.60 (s, 9H), 1.46-1.39 (m, 9H); ES-LCMS m/z 643.3 [M+H]⁺.

Step 3:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-tetrahydrofuran-2-yl-pyrazolo[1,5-a]pyrimidin-7-amine(I-226)

To a stirred solution of tert-butyl3-[2-[tert-butoxycarbonyl-[5-(5-fluoro-3-pyridyl)-3-tetrahydrofuran-2-yl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(70 mg, 100.96 μmol, 1 eq) in DCM (8 mL) was added ZnBr₂ (2.27 g, 10.10mmol, 100 eq). The reaction mixture was stirred at 25° C. for 15 h. TLC(PE/EtOAc=1/1, R_(f)=0.35) indicated one major new spot was detected.MeOH (30 mL) was added into the reaction mixture and diluted with water(50 mL), adjusted pH to 8-9 with saturated NaHCO₃ solution, extractedwith EtOAc (30 mL×3). The combined organic layers were dried overNa₂SO₄, filtered and concentrated under reduced pressure to yield aresidue which was purified by flash silica gel chromatography (fromPE/EtOAc=100/1 to 2/1, TLC: PE/EtOAc=1/1, R_(f)=0.35) to yield a residuewhich was re-purified by preparative HPLC (basic condition; column:Gemini 150*25 5 u; mobile phase: [water (0.05% ammonia hydroxidev/v)-ACN]; B %: 55%-75%, 8 min). The desired fraction was lyophilized toyield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-tetrahydrofuran-2-yl-pyrazolo[1,5-a]pyrimidin-7-amine(5.91 mg, 12.85 μmol, 12.7% yield, 96.2% purity) as a white solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.80 (s, 1H), 8.50 (s, 1H), 8.03 (s, 2H),7.85 (d, J=9.5 Hz, 1H), 7.69 (d, J=7.5 Hz, 1H), 7.36 (d, J=7.7 Hz, 1H),7.26-7.18 (m, 2H), 7.05 (d, J=2.0 Hz, 1H), 6.54 (s, 1H), 6.07 (s, 1H),5.29 (t, J=7.3 Hz, 1H), 4.12 (q, J=7.3 Hz, 1H), 3.99-3.90 (m, 1H), 3.85(q, J=6.3 Hz, 2H), 3.25 (t, J=6.5 Hz, 2H), 2.39-2.28 (m, 2H), 2.24-2.15(m, 1H), 2.14-2.04 (m, 1H); ES-LCMS m/z 443.2 [M+H]⁺.

Example 177

Synthesis of I-227

Synthetic Scheme:

Step 1: 1,3-Benzodioxol-4-yl(trimethyl)stannane

To a solution of 4-iodo-1,3-benzodioxole (100 mg, 403.20 μmol, 1 eq) in1,4-dioxane (5 mL) was added trimethyl(trimethylstannyl)stannane (132.10mg, 403.20 μmol, 83.61 μL, 1 eq) and4-diphenylphosphanylbutyl(diphenyl)phosphane (17.20 mg, 40.32 μmol, 0.1eq) was purged with N₂ for 15 min. Pd(OAc)₂ (9.05 mg, 40.32 μmol, 0.1eq) was added and the mixture was stirred at 110° C. for 3 h. TLC(PE/EtOAc=10/1, R_(f)=0.44) indicated one major new spot with lowerpolarity was detected. The reaction mixture was concentrated underreduced pressure to yield a residue which was purified on silica gelcolumn chromatography (from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1,R_(f)=0.44) to yield the product 1,3-benzodioxol-4-yl(trimethyl)stannane(80 mg, 252.70 μmol, 62.7% yield, 90.0% purity) as colorless oil. ¹H NMR(400 MHz, CDCl₃) δ ppm 6.88-6.76 (m, 3H), 5.91 (s, 2H), 0.42-0.26 (s,9H).

Step 2: tert-Butyl3-[2-[[5-(1,3-benzodioxol-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate

To a solution of 1,3-benzodioxol-4-yl(trimethyl)stannane (50 mg, 157.94μmol, 1 eq) and tert-butyl3-[2-[tert-butoxycarbonyl-(5-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate (75.06 mg, 134.25 μmol, 0.85 eq) in DMF(5 mL) was purged with N₂ for 25 min. Pd(dppf)Cl₂ (10.98 mg, 15.00 μmol,0.095 eq) and CuI (2.86 mg, 15.00 μmol, 0.095 eq) was added and themixture was stirred at 110° C. for 2 h under microwave. The reactionmixture was filtered and concentrated under reduced pressure to yield aresidue which was purified by preparative TLC (TLC: PE/EtOAc=10/1,R_(f)=0.46) to yield the product tert-butyl3-[2-[[5-(1,3-benzodioxol-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate(80 mg, 106.42 μmol, 67.4% yield, 85.1% purity) as yellow oil. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.03 (s, 2H), 7.99 (s, 1H), 7.87 (d, J=7.7 Hz,1H), 7.50 (d, J=7.1 Hz, 1H), 7.39 (br s, 2H), 7.23-7.14 (m, 1H),7.03-6.96 (m, 1H), 6.95-6.90 (m, 1H), 6.09-6.04 (m, 2H), 4.19 (t, J=7.5Hz, 2H), 3.46-3.37 (m, 1H), 3.06 (t, J=6.9 Hz, 2H), 1.63 (s, 9H), 1.45(d, J=6.4 Hz, 6H), 1.36 (s, 9H); ES-LCMS m/z 640.4 [M+H]⁺.

Step 3:5-(1,3-Benzodioxol-4-yl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-227)

A solution of tert-butyl3-[2-[[5-(1,3-benzodioxol-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]-tert-butoxycarbonyl-amino]ethyl]indole-1-carboxylate(80 mg, 106.42 μmol, 1 eq) in HCl/MeOH (4 M, 5 mL, 187.94 eq) was wasstirred at 25° C. for 12 h. LC-MS showed the starting material was notconsumed completely. The mixture was stirred at 25° C. for another 12 h.The reaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: PhenomenexGemini 150*25 mm*10 um; mobile phase: [water (0.05% HCl)-ACN]; B %:35%-65%, 9 min) followed by lyophilization to yield5-(1,3-benzodioxol-4-yl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(47.73 mg, 100.08 μmol, 94.0% yield, 99.8% purity, HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.16 (s, 1H), 7.51 (d, J=7.8 Hz,1H), 7.14 (d, J=8.0 Hz, 1H), 7.05-7.00 (m, 2H), 6.97-6.92 (m, 2H),6.86-6.80 (m, 1H), 6.53 (d, J=8.0 Hz, 1H), 6.07 (s, 2H), 5.76 (s, 1H),3.96 (t, J 5.8 Hz, 2H), 3.23-3.18 (m, 3H), 1.33 (d, J=6.8 Hz, 6H);ES-LCMS m/z 440.3 [M+H]⁺.

Example 178

Synthesis of I-229

Synthetic Scheme:

Step 1: tert-Butyl(3R)-3-[tert-butoxycarbonyl-(3-isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

tert-Butyl(3R)-3-[tert-butoxycarbonyl-(5-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate (200 mg, 316.66 μmol, 1eq), pyrimidin-5-ylboronic acid (47.08 mg, 379.99 μmol, 1.2 eq), Cs₂CO₃(309.52 mg, 949.98 μmol, 3 eq) and Pd(dppf)Cl₂ (23.17 mg, 31.67 μmol,0.1 eq) were taken up into a microwave tube in 1,4-dioxane (6 mL) andH₂O (2 mL). The sealed tube was heated at 110° C. for 1 h undermicrowave (2 bar). The mixture was concentrated and then saturatedNaHCO₃ (10 mL) was added, extracted with EtOAc (10 mL×3). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated toyield a residue which was purified on silica gel column chromatography(from PE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1, R_(f)=0.30) to yieldtert-butyl(3R)-3-[tert-butoxycarbonyl-(3-isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate (180 mg,288.58 μmol, 91.1% yield, 100.0% purity) as a yellow gum. ¹H NMR (400MHz, CD₃OD) δ ppm 9.39 (s, 2H), 9.22 (s, 1H), 8.14 (s, 1H), 7.94 (d,J=7.9 Hz, 1H), 7.46 (s, 1H), 7.19 (br s, 1H), 7.14-7.02 (m, 2H), 4.79(m, 1H), 3.48 (m, 1H), 3.43-3.36 (m, 2H), 3.18-3.08 (m, 2H), 2.36-2.28(m, 2H), 1.63 (s, 9H), 1.45 (d, J=7.1 Hz, 6H), 1.30 (s, 9H); ES-LCMS m/z624.4 [M+H]⁺.

Step 2:(3R)—N-(3-Isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-229)

To a solution of tert-butyl(3R)-3-[tert-butoxycarbonyl-(3-isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(180 mg, 288.58 μmol, 1 eq) in DCM (5 mL) was added TFA (1 mL). Themixture was degassed and purged with N₂ for 3 times, the mixture wasstirred under N₂ atmosphere at 20° C. for 3 h. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 μm;mobile phase: [water (0.05% HCl)-ACN]; B %: 50%-80%, 9 min) to yield theresidue which was purified by preparative HPLC (column: PhenomenexGemini 150*25 mm*10 μm; mobile phase: [water (0.05% ammonia hydroxidev/v)-ACN]; B %: 56%-86%, 10 min) followed by lyophilization to yield(3R)—N-(3-isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(12 mg, 28.33 μmol, 9.8% yield, 100.0% purity) as a yellow solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 9.51 (s, 2H), 9.21 (s, 1H), 7.95 (s, 1H), 7.39(d, J=7.7 Hz, 1H), 7.27 (d, J=8.2 Hz, 1H), 7.07-7.01 (m, 1H), 6.99-6.93(m, 1H), 6.84 (s, 1H), 4.47 (m, J=8.6 Hz, 1H), 3.37-3.34 (m, 2H),3.14-3.03 (m, 1H), 3.00-2.88 (m, 2H), 2.41-2.33 (m, 1H), 2.28-2.19 (m,1H), 1.42 (d, J=6.8 Hz, 6H); ES-LCMS m/z 424.2 [M+H]⁺.

Example 179

Synthesis of I-230

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-vinyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-230)

A mixture of5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-iodo-pyrazolo[1,5-a]pyrimidin-7-amine(55 mg, 109.71 μmol, 1 eq),4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (50.69 mg, 329.14 μmol,55.83 μL, 3 eq), Cs₂CO₃ (178.74 mg, 548.57 μmol, 5 eq) and Pd(dppf)Cl₂(40.14 mg, 54.86 μmol, 0.5 eq) in 1,4-dioxane (3 mL) and H₂O (1.5 mL)was bubbled with N₂ for 2 minutes and sealed. The reaction mixture wasirradiated under microwave (2 bar) at 120° C. for 2 h. The reactionmixture was diluted with H₂O (10 mL) and extracted with EtOAc (10 mL×3).The organic layer was dried over Na₂SO₄, filtered and concentrated underreduced pressure to yield a residue which was purified by preparativeTLC (PE/EtOAc=2/1, R_(f)=0.23) and then by preparative HPLC (column:Gemini 150*25 5 u; mobile phase: [water (0.05% ammonia hydroxidev/v)-ACN]; B %: 60%-90%, 8 min). The desired fraction was lyophilized toyield5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-vinyl-pyrazolo[1,5-a]pyrimidin-7-amine(15.4 mg, 38.65 μmol, 35.2% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.82 (s, 1H), 8.51 (d, J=2.4 Hz, 1H), 8.06(s, 2H), 7.95-7.88 (m, 1H), 7.69 (d, J=7.6 Hz, 1H), 7.39-7.33 (m, 1H),7.27-7.18 (m, 2H), 7.06 (d, J=2.0 Hz, 1H), 6.90 (dd, J=11.2, 17.6 Hz,1H), 6.55 (t, J=6.4 Hz, 1H), 6.11 (s, 1H), 6.07 (dd, J=1.6, 17.6 Hz,1H), 5.27 (dd, J=2.0, 11.2 Hz, 1H), 3.85 (q, J=6.4 Hz, 2H), 3.27 (t,J=6.4 Hz, 2H); ES-LCMS m/z 399.2 [M+H]⁺.

Example 180

Synthesis of I-232

Synthetic Scheme:

Step1:5-(5-Fluoro-3-pyridyl)-N-[(2R)-5-methoxyindan-2-yl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine

To a mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidine (50mg, 190.35 μmol, 1 eq), (2R)-5-methoxyindan-2-amine (38.01 mg, 190.35μmol, 1 eq, HCl) in i-PrOH (5 mL) was added DIEA (73.80 mg, 571.06 μmol,99.47 uL, 3 eq). The mixture was stirred at 80° C. for 12 h. The mixturewas concentrated to yield a crude compound5-(5-fluoro-3-pyridyl)-N-[(2R)-5-methoxyindan-2-yl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(70 mg, 130.90 μmol, 68.8% yield, 72.8% purity) as yellow oil which wasused in the next step without further purification; ES-LCMS m/z390.2[M+H]⁺.

Step 2:(2R)-2-[[5-(5-Fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(I-232)

A mixture of5-(5-fluoro-3-pyridyl)-N-[(2R)-5-methoxyindan-2-yl]-3-methyl-pyrazolo[1,5-a]pyrimidin-7-amine(70 mg, 130.90 μmol, 1 eq) in HBr solution (7.45 g, 55.25 mmol, 5 mL,60%, 422.06 eq) was stirred at 80° C. for 1 h. The mixture wasconcentrated to yield a residue was purified by preparative HPLC(column: Phenomenex Gemini 150*25 mm*10 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 30%-60%, 6 min). The desired fraction was lyophilized toyield(2R)-2-[[5-(5-fluoro-3-pyridyl)-3-methyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(17.22 mg, 38.27 mol, 29.2% yield, 99.6% purity, 2HCl, ee=98.2%) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.98 (s, 1H), 8.80 (d, J=2.4Hz, 1H), 8.29 (td, J=2.4, 9.2 Hz, 1H), 8.11 (s, 1H), 7.06 (d, J=8.0 Hz,1H), 6.94 (s, 1H), 6.69 (s, 1H), 6.63 (dd, J=2.4, 8.0 Hz, 1H), 5.07-4.96(m, 1H), 3.47-3.39 (m, 2H), 3.23-3.15 (m, 2H), 2.36 (s, 3H); ES-LCMS m/z376.1 [M+H]⁺.

Example 181

Synthesis of I-234

Synthetic Scheme:

Step 1:5-(5-Fluoro-3-pyridyl)-3-isopropyl-N-[(2R)-5-methoxyindan-2-yl]pyrazolo[1,5-a]pyrimidin-7-amine

To a mixture of7-chloro-5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidine(50 mg, 154.79 μmol, 1 eq), (2R)-5-methoxyindan-2-amine (27.79 mg,170.27 μmol, 1.1 eq) in i-PrOH (5 mL) was added DIEA (60.01 mg, 464.36mol, 80.88 μL, 3 eq). The mixture was stirred at 80° C. for 12 h. Themixture was concentrated to yield the product of5-(5-fluoro-3-pyridyl)-3-isopropyl-N-[(2R)-5-methoxyindan-2-yl]pyrazolo[1,5-a]pyrimidin-7-amine(60 mg, 102.65 μmol, 66.3% yield, 71.4% purity) as yellow oil which wasused in the next step without further purification. ES-LCMS m/z418.2[M+H]⁺.

Step 2:(2R)-2-[[5-(5-Fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol(I-234)

A mixture of5-(5-fluoro-3-pyridyl)-3-isopropyl-N-[(2R)-5-methoxyindan-2-yl]pyrazolo[1,5-a]pyrimidin-7-amine (60 mg, 102.04 μmol, 1 eq) in HBr solution(5.29 g, 39.22 mmol, 3.55 mL, 60%, 384.39 eq) was stirred at 120° C. for2 h. The mixture was concentrated to yield a residue which was purifiedby preparative HPLC (column: Phenomenex Gemini 150*25 mm*10 um; mobilephase: [water (0.05% HCl)-ACN]; B %: 45%-75%, 9 min). The desiredfraction was lyophilized to yield(2R)-2-[[5-(5-fluoro-3-pyridyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]indan-5-ol (13.92 mg, 31.34 μmol, 30.7% yield,99.0% purity, HCl) (97.1% ee) as a yellow solid. ¹H NMR (400 MHz, CD₃OD)δ ppm 8.98 (s, 1H), 8.80 (d, J=2.0 Hz, 1H), 8.32-8.29 (m, 1H), 8.21 (s,1H), 7.05 (d, J=8.0 Hz, 1H), 6.92 (s, 1H), 6.62 (dd, J=2.4, 8.4 Hz, 1H),6.61 (m, 1H), 5.02-4.93 (m, 1H), 3.49-3.40 (m, 2H), 3.37-3.31 (m, 1H),3.21-3.09 (m, 2H), 1.36 (d, J=6.8 Hz, 6H); ES-LCMS m/z 404.3 [M+H]⁺.

Example 182

Synthesis of I-235

Synthetic Scheme:

Step 1:N-[2-(7-Fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-7-amine(I-235)

To a solution of7-chloro-5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidine(30 mg, 87.92 μmol, 1 eq) and2-(7-fluoro-2-methyl-1H-indol-3-yl)ethanamine (24.15 mg, 105.51 μmol,1.2 eq) in i-PrOH (5 mL) was added DIEA (34.09 mg, 263.77 μmol, 45.94μL, 3.0 eq). The mixture was stirred at 80° C. for 9 h. The reactionmixture was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Phenomenex Gemini 150*25 mm*10um; mobile phase: [water (0.05% HCl)-ACN]; B %: 50%-80%, 10 min)followed by lyophilization to yieldN-[2-(7-fluoro-2-methyl-1H-indol-3-yl)ethyl]-5-(5-fluoro-3-pyridyl)-3-(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-7-amine(10.90 mg, 19.97 μmol, 22.7% yield, 99.9% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.86 (dd, J=1.3, 2.5 Hz, 1H), 8.74(d, J=1.5 Hz, 1H), 8.34 (s, 1H), 7.94 (td, J=2.3, 8.7 Hz, 1H), 7.42 (d,J=8.0 Hz, 1H), 6.98 (dt, J=4.8, 7.9 Hz, 1H), 6.67 (dd, J=7.9, 11.4 Hz,1H), 5.92 (s, 1H), 3.98-3.91 (m, 2H), 3.21-3.14 (m, 2H), 2.13-2.05 (m,3H); ES-LCMS m/z 473.2 [M+H]⁺.

Example 183

Synthesis of I-237

Synthetic Scheme:

Step 1:3-Ethyl-5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(I-237)

A mixture of5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]-3-vinyl-pyrazolo[1,5-a]pyrimidin-7-amine(140 mg, 196.77 μmol, 1 eq) and Pd/C (50 mg, 10% purity) in EtOAc (20mL) was stirred under H₂ (15 Psi) at 25° C. for 12 h. The reactionmixture was filtered. The filtrate was concentrated under reducedpressure to yield a residue which was purified by preparative HPLC(column: Phenomenex Gemini C18 250*50 mm*10 μm; mobile phase: [water(0.05% HCl)-ACN]; B %: 35%-65%, 10 min). The desired fraction waslyophilized to yield3-ethyl-5-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrazolo[1,5-a]pyrimidin-7-amine(7.13 mg, 15.06 μmol, 7.7% yield, 100.0% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.65 (d, J=2.4 Hz, 1H), 8.37 (s,1H), 8.11 (s, 1H), 7.53 (d, J=8.0 Hz, 1H), 7.50-7.46 (m, 1H), 7.14 (d,J=8.0 Hz, 1H), 6.99 (s, 1H), 6.96 (t, J=7.6 Hz, 1H), 6.87-6.82 (m, 1H),5.78 (s, 1H), 3.98 (t, J=5.6 Hz, 2H), 3.20 (t, J=5.6 Hz, 2H), 2.72 (q,J=7.6 Hz, 2H), 1.28 (t, J=7.6 Hz, 3H); ES-LCMS m/z 401.2 [M+H]⁺.

Example 184

Synthesis of I-240

Synthetic Scheme:

Step 1: tert-Butyl3-[2-[tert-butoxycarbonyl-[3-isopropyl-5-(2-methoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate

To a solution of tert-butyl3-[2-[tert-butoxycarbonyl-(5-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate(100 mg, 178.86 μmol, 1 eq) in 1,4-dioxane (6 mL) and water (2 mL) wasadded (2-methoxyphenyl)boronic acid (81.53 mg, 536.57 μmol, 3 eq), K₃PO₄(113.89 mg, 536.57 μmol, 3 eq) and sphos palladacycle (13.61 mg, 17.89μmol, 0.1 eq). The mixture was stirred at 110° C. for 1 h under N₂atmosphere. The mixture was concentrated under reduced pressure, thenwater (30 mL) was added. The mixture was extracted with EtOAc (50 mL×3).The combined organic layers were washed with brine (50 mL), dried overNa₂SO₄, filtered and concentrated to yield tert-butyl3-[2-[tert-butoxycarbonyl-[3-isopropyl-5-(2-methoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(200 mg, 164.28 μmol, 91.8% yield, 51.4% purity) as yellow oil which wasused in the next step without further purification. ¹H NMR (400 MHz,CD₃OD) δ ppm 7.96 (s, 1H), 7.92-7.87 (m, 1H), 7.73 (dd, J=1.7, 7.6 Hz,1H), 7.65-7.60 (m, 1H), 7.47 (dd, J=1.4, 8.7 Hz, 1H), 7.19-7.13 (m, 2H),7.07 (s, 1H), 6.97-6.94 (m, 1H), 6.94-6.91 (m, 2H), 4.1 (m, 2H), 3.81(s, 3H), 3.36 (s, 1H), 3.06 (t, J=6.4 Hz, 2H), 1.61-1.55 (m, 9H),1.48-1.45 (m, 6H), 1.37-1.35 (m, 9H); ES-LCMS m/z 626.4 [M+H]⁺.

Step 2:N-[2-(1H-Indol-3-yl)ethyl]-3-isopropyl-5-(2-methoxyphenyl)pyrazolo[1,5-a] pyrimidin-7-amine (I-240

To a solution of tert-butyl3-[2-[tert-butoxycarbonyl-[3-isopropyl-5-(2-methoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate (185 mg,151.96 μmol, 1 eq) in DCM (10 mL) was added TFA (3.08 g, 27.01 mmol, 2mL, 177.76 eq). The mixture was stirred at 25° C. for 2 h under N₂atmosphere. The reaction mixture was concentrated under reduced pressureto yield a residue which was purified by preparative HPLC (column:Phenomenex Gemini 150*25 mm*10 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 23%-53%, 10 min) followed by lyophilization to yieldN-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-5-(2-methoxyphenyl)pyrazolo[1,5-a]pyrimidin-7-amine(27.57 mg, 63.69 μmol, 41.9% yield, 98.3% purity) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 8.11 (s, 1H), 7.57-7.47 (m, 2H), 7.25 (d,J=8.0 Hz, 1H), 7.13 (d, J=8.3 Hz, 1H), 7.06-6.95 (m, 3H), 6.84 (t, J=7.5Hz, 2H), 5.79 (s, 1H), 3.93 (t, J=6.1 Hz, 2H), 3.83 (s, 3H), 3.21 (t,J=6.1 Hz, 2H), 3.19-3.12 (m, 1H), 1.33 (d, J=6.8 Hz, 6H); ES-LCMS m/z426.2 [M+H]⁺.

Example 185

Synthesis of I-241

Synthetic Scheme:

Step 1: tert-Butyl3-[2-[tert-butoxycarbonyl-(3-isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate

Tert-butyl3-[2-[tert-butoxycarbonyl-(5-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate (100 mg, 178.86 μmol, 1 eq),pyrimidin-5-ylboronic acid (22.16 mg, 178.86 μmol, 1 eq), Pd(dppf)Cl₂(13.09 mg, 17.89 μmol, 0.1 eq) and Cs₂CO₃ (174.82 mg, 536.57 μmol, 3 eq)were taken up into a microwave tube in H₂O (2 mL) and 1,4-dioxane (6mL). The sealed tube was heated at 110° C. for 1 h under microwave. Themixture was concentrated and saturated NaHCO₃ (10 mL) was added,extracted with EtOAc (10 mL×3). The combined organic layers were driedover Na₂SO₄, filtered and concentrated to yield tert-butyl3-[2-[tert-butoxycarbonyl-(3-isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate(100 mg, 167.31 μmol, 93.5% yield, crude) as brown solid which was usedin the next step without further purification. ¹H NMR (400 MHz, CD₃OD) δppm 9.23-9.22 (m, 1H), 9.21-9.20 (m, 1H), 8.97 (s, 1H), 7.81 (d, J=7.6Hz, 1H), 7.45 (d, J=7.6 Hz, 2H), 7.41 (d, J=8.4 Hz, 1H), 7.09-7.02 (m,2H), 6.93 (t, J=8.0 Hz, 1H), 4.33 (d, J=6.4 Hz, 2H), 4.30 (s, 1H), 3.11(t, J=6.4 Hz, 2H), 1.59 (s, 9H), 1.39 (s, 6H), 1.37 (s, 9H); ES-LCMS m/z598.4 [M+H]⁺.

Step 2:N-[2-(1H-Indol-3-yl)ethyl]-3-isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-amine(I-241)

To a solution of tert-butyl3-[2-[tert-butoxycarbonyl-(3-isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate(100 mg, 167.31 μmol, 1 eq) in DCM (7.5 mL) was added TFA (1.5 mL). Themixture was degassed and purged with N₂ for 3 times and stirred under N₂atmosphere at 20° C. for 3 h. The reaction mixture was concentratedunder reduced pressure to give a residue which was purified bypreparative HPLC (column: Phenomenex Gemini C18 250*50 mm*10 μm; mobilephase: [water (0.05% HCl)-ACN]; B %: 35%-65%, 10 min). The desiredfraction was lyophilized to yieldN-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-5-pyrimidin-5-yl-pyrazolo[1,5-a]pyrimidin-7-amine(17.94 mg, 34.12 μmol, 20.4% yield, 96.4% purity, 3HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.31 (s, 1H), 8.68 (s, 2H), 8.18(s, 1H), 7.50 (d, J=7.9 Hz, 1H), 7.16 (d, J=8.0 Hz, 1H), 7.03 (s, 1H),6.99-6.92 (m, 1H), 6.83-6.77 (m, 1H), 5.83 (s, 1H), 4.04-3.97 (m, 2H),3.25-3.16 (m, 3H), 1.35 (d, J=6.8 Hz, 6H); ES-LCMS m/z 398.1 [M+H]⁺.

Example 186

Synthesis of I-242

Synthetic Scheme:

Step 1: tert-Butyl N-(2-bromo-4,6-difluoro-phenyl)carbamate

tert-Butyl3-[2-[tert-butoxycarbonyl-(5-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate (100 mg, 178.86 μmol, 1 eq),(3-fluorophenyl)boronic acid (75.08 mg, 536.57 μmol, 3 eq), Pd(dppf)Cl₂(13.09 mg, 17.89 μmol, 0.1 eq), Cs₂CO₃ (174.82 mg, 536.57 μmol, 3 eq)and H₂O (2 mL) were taken up into a microwave tube in 1,4-dioxane (6mL). The sealed tube was heated at 110° C. for 1 h under microwave. Thereaction mixture was concentrated under reduced pressure to give aresidue which was diluted with aq. NaHCO₃ (20 mL) and extracted withEtOAc (50 mL×3). The combined organic layers were dried over Na₂SO₄,filtered and concentrated under reduced pressure to yield tert-butyl3-[2-[tert-butoxycarbonyl-[5-(3-fluorophenyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate(100 mg, 97.77 μmol, 54.7% yield, 60.0% purity) as a brown solid whichwas used in the next step without further purification. ¹H NMR (400 MHz,CD₃OD) δ ppm 7.98 (s, 1H), 7.94 (d, J=7.6 Hz, 1H), 7.59 (d, J=1.2 Hz,1H), 7.56 (d, J=2.4 Hz, 1H), 7.53 (s, 1H), 7.51 (s, 1H), 7.48 (d, J=3.6Hz, 1H), 7.46 (d, J=2.0 Hz, 1H), 7.26 (s, 1H), 7.19-7.15 (m, 1H),6.65-6.57 (m, 1H), 4.24-4.21 (m, 2H), 3.58-3.44 (m, 1H), 3.11 (t, J=6.4Hz, 2H), 1.56 (s, 9H), 1.50-1.45 (m, 6H), 1.37 (s, 9H); ES-LCMS m/z614.3 [M+H]⁺.

Step 2:5-(3-Fluorophenyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine (I-242)

A solution of tert-butyl3-[2-[tert-butoxycarbonyl-[5-(3-fluorophenyl)-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl]amino]ethyl]indole-1-carboxylate (80 mg, 78.21μmol, 1 eq) in HCl/MeOH (4 M, 3 mL, 153.43 eq) was stirred at 25° C. for2 h under N₂ atmosphere. The reaction mixture was concentrated underreduced pressure to give a residue which was purified by preparativeHPLC (column: Phenomenex Gemini C18 250*50 mm*10 um; mobile phase:[water (0.05% HCl)-ACN]; B %: 35%-65%, 10 min) followed bylyophilization to yield5-(3-fluorophenyl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(38.32 mg, 76.97 μmol, 98.4% yield, 97.7% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.16 (s, 1H), 7.55-7.46 (m, 2H),7.31-7.36 (m, 1H), 7.20 (d, J=8.0 Hz, 1H), 7.10 (d, J=8.8 Hz, 1H),7.06-6.92 (m, 3H), 6.85 (t, J=7.6 Hz, 1H), 5.74 (s, 1H), 4.08-3.92 (m,2H), 3.29-3.24 (m, 1H), 3.24-3.19 (m, 2H), 1.34 (d, J=6.8 Hz, 6H);ES-LCMS m/z 414.2 [M+H]⁺.

Example 187

Synthesis of I-245

Synthetic Scheme:

Step 1: 3-Isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol

A mixture of methyl 3-oxo-3-(3-pyridyl)propanoate (440 mg, 2.46 mmol, 1eq) and 4-isopropyl-1H-pyrazol-5-amine (307.39 mg, 2.46 mmol, 1 eq) inAcOH (20 mL) was stirred at 120° C. for 0.5 h. The reaction mixture wasconcentrated under reduced pressure to yield3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol (600 mg, 1.60mmol, 65.3% yield, crude, 2HOAC) as brown oil which was used in the nextstep without further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm9.19-9.17 (m, 1H), 8.83-8.78 (m, 1H), 8.27 (td, J=1.9, 8.0 Hz, 1H),7.50-7.42 (m, 2H), 7.14 (s, 1H), 2.84-2.75 (m, 1H), 1.22 (d, J=2.0 Hz,6H); ES-LCMS m/z 255.1 [M+H]⁺.

Step 2: 7-Chloro-3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidine

A solution of 3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-ol(600 mg, 1.60 mmol, 1 eq, 2HOAC) in POCl₃ (15 mL) was degassed andpurged with N₂ for 3 times. The mixture was stirred at 110° C. for 3 h.The reaction mixture was concentrated under reduced pressure to yield aresidue which adjusted to pH to 7-8 with TEA. The residue was purifiedon silica gel column chromatography (from PE/EtOAc=1/0 to 10/3, TLC:PE/EtOAc=3/1, R_(f)=0.35) to yield7-chloro-3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidine (100 mg,322.66 μmol, 20.1% yield, 88.0% purity) as a green solid. ¹H NMR (400MHz, CDCl₃) δ ppm 9.33 (d, J=1.8 Hz, 1H), 8.74 (dd, J=1.5, 4.9 Hz, 1H),8.46 (td, J=2.0, 7.9 Hz, 1H), 8.13 (s, 1H), 7.50 (dd, J=4.7, 8.0 Hz,1H), 7.40 (s, 1H), 3.42 (td, J=6.9, 13.8 Hz, 1H), 1.46 (d, J=7.1 Hz,6H); ES-LCMS m/z 273.0 [M+H]⁺.

Step 3:(3R)—N-[3-Isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-245)

A mixture of 7-chloro-3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidine(60 mg, 193.60 μmol, 1 eq), (3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(36.06 mg, 193.60 μmol, 1 eq) and DIEA (75.06 mg, 580.79 μmol, 101.16μL, 3 eq) in i-PrOH (10 mL) was degassed and purged with N₂ for 3 times.The mixture was stirred at 80° C. for 3 h under N₂ atmosphere. Thereaction mixture was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: PhenomenexGemini 150*25 mm*10 μm; mobile phase: [water (0.05% HCl)-ACN]; B %:42%-70%, 7 min) followed by lyophilization to yield(3R)—N-[3-isopropyl-5-(3-pyridyl)pyrazolo[1,5-a]pyrimidin-7-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(40.65 mg, 76.42 μmol, 39.5% yield, 100.0% purity, 3HCl) as a red solid.¹H NMR (400 MHz, CD₃OD) δ ppm 9.52 (s, 1H), 9.17 (J=8.2 Hz, 1H), 8.97(J=5.3 Hz, 1H), 8.21-8.15 (m, 2H), 7.34 (d, J=7.7 Hz, 1H), 7.23 (d,J=7.9 Hz, 1H), 7.10 (s, 1H), 7.00 (t, J=7.2 Hz, 1H), 6.95-6.88 (m, 1H),4.62 (br s, 1H), 3.41 (td, J=7.0, 13.8 Hz, 1H), 3.27 (J=4.9 Hz, 1H),3.13 (J=9.7 Hz, 1H), 3.01-2.91 (m, 2H), 2.42-2.22 (m, 2H), 1.42 (dd,J=3.1, 6.8 Hz, 6H); ES-LCMS m/z 423.2 [M+H]⁺.

Example 188

Synthesis of I-246

Synthetic Scheme:

Step 1: 3-Isopropyl-4H-pyrazolo[1,5-a]pyrimidine-5,7-dione

Na (367.33 mg, 15.98 mmol, 2 eq) was added into EtOH 25 mL and themixture was stirred at 24° C. for 2 h. 4-Isopropyl-1H-pyrazol-5-amine (1g, 7.99 mmol, 1 eq) and diethyl propanedioate (1.41 g, 8.79 mmol, 1.33mL, 1.1 eq) were added into the above reaction mixture and stirred at80° C. for 5 h. The solution was cooled to room temperature andfiltered. The obtained solid was washed with EtOH and dissolved in water(20 mL). Then the solution was acidified to pH=1-2 with concentratedhydrochloric acid in an ice-bath. The precipitate was filtered, washedwith water and dried under reduced pressure to yield3-isopropyl-4H-pyrazolo[1,5-a]pyrimidine-5,7-dione (300 mg, 1.55 mmol,19.4% yield, 100.0% purity) as a yellow solid which was used in the nextstep without further purification. ¹H NMR (400 MHz, CD₃OD) δ ppm7.80-7.73 (m, 2H), 3.02 (td, J=6.7, 13.7 Hz, 1H), 1.28 (d, J=7.1 Hz,6H); ES-LCMS m/z 194.2 [M+H]⁺.

Step 2: 5,7-Dichloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine

3-Isopropyl-4H-pyrazolo[1,5-a]pyrimidine-5,7-dione (285 mg, 1.48 mmol, 1eq) was added into POCl₃ (20 mL). The reaction mixture was stirred at110° C. for 12 h. The reaction mixture was concentrated under reducedpressure to give a residue which was purified by flash silica gelchromatography (from PE/EtOAc=100/1 to 5/1, TLC: PE/EtOAc=10/1,R_(f)=0.30) to yield 5,7-dichloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine(300 mg, 1.30 mmol, 88.4% yield, 100.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.10 (s, 1H), 6.94-6.90 (m, 1H), 3.37-3.26(m, 1H), 1.37 (d, J=6.8 Hz, 6H); ES-LCMS m/z 230.1, 232.1 [M+H]⁺.

Step 3:5-Chloro-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine

To a solution of 5,7-dichloro-3-isopropyl-pyrazolo[1,5-a]pyrimidine (200mg, 869.21 μmol, 1 eq) in i-PrOH (6 mL) was added DIEA (337.02 mg, 2.61mmol, 454.20 μL, 3 eq) and 2-(1H-indol-3-yl)ethanamine (167.11 mg, 1.04mmol, 1.2 eq). The mixture was stirred at 90° C. for 12 h. The reactionmixture was concentrated under reduced pressure to give a residue whichwas purified by flash silica gel chromatography (from PE/EtOAc=100/1 to3/1, TLC: PE/EtOAc=3/1, R_(f)=0.60) to yield5-chloro-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(240 mg, 672.83 μmol, 77.4% yield, 99.2% purity) as yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.11 (s, 1H), 7.81 (s, 1H), 7.62 (d, J=7.8 Hz,1H), 7.41 (d, J=8.3 Hz, 1H), 7.27-7.22 (m, 1H), 7.20-7.13 (m, 1H), 7.11(d, J=2.3 Hz, 1H), 6.45 (s, 1H), 5.88 (s, 1H), 3.71 (q, J=6.6 Hz, 2H),3.33-3.18 (m, 3H), 1.33 (d, J=7.0 Hz, 6H); ES-LCMS m/z 354.1, 355.1[M+H]⁺.

Step 4: tert-Butyl3-[2-[tert-butoxycarbonyl-(5-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate

To a solution of5-chloro-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(120 mg, 336.42 μmol, 1 eq) in 1,4-dioxane (6 mL) was added DMAP (123.30mg, 1.01 mmol, 3 eq) and (Boc)₂O (183.56 mg, 841.04 μmol, 193.22 μL, 2.5eq). The mixture was stirred at 110° C. for 5 h. The reaction mixturewas concentrated under reduced pressure to give a residue which waspurified by flash silica gel chromatography (from PE/EtOAc=100/1 to 3/1,TLC: PE/EtOAc=3/1, R_(f)=0.75) to yield tert-butyl3-[2-[tert-butoxycarbonyl-(5-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate(150 mg, 262.33 μmol, 77.9% yield, 96.9% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.09 (d, J=6.4 Hz, 1H), 7.97 (s, 1H), 7.43(d, J=7.9 Hz, 1H), 7.35 (s, 1H), 7.32-7.27 (m, 1H), 7.20-7.13 (m, 1H),6.39 (s, 1H), 4.21-4.15 (m, 2H), 3.28 (td, J=7.0, 13.9 Hz, 1H), 3.06 (t,J=7.4 Hz, 2H), 1.67 (s, 9H), 1.41 (s, 9H), 1.36 (d, J=7.1 Hz, 6H);ES-LCMS m/z 554.3, 556.3 [M+H]⁺.

Step 5:5-(1,3-Benzodioxol-5-yl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(I-246)

To a solution of tert-butyl3-[2-[tert-butoxycarbonyl-(5-chloro-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-yl)amino]ethyl]indole-1-carboxylate(100 mg, 174.88 μmol, 1 eq) in 1,4-dioxane (6 mL) and water (2 mL) wasadded K₃PO₄ (111.37 mg, 524.65 μmol, 3 eq), sphos palladacycle (13.30mg, 17.49 μmol, 0.1 eq) and 1,3-benzodioxol-5-ylboronic acid (87.06 mg,524.65 μmol, 3 eq). The mixture was degassed and purged with N₂ threetimes and stirred at 110° C. for 1 h under microwave. The reactionmixture was diluted with EtOAc (50 mL) and filtered through a pad ofcelite. The filtrate was concentrated under reduced pressure to give aresidue which was dissolved in DCM (1 mL) and HCl/EtOAc (1 mL, 4M) wasadded. The reaction mixture was stirred at 25° C. for 14 h. The reactionmixture was diluted with water (50 mL) and adjusted to pH=2 by 10% aq.NaOH solution. The mixture was extracted with EtOAc (50 mL×3), driedover anhydrous Na₂SO₄, filtered and the filtrate was concentrated togive the residue which was purified by preparative HPLC (HCl condition;column: Phenomenex Gemini 150*25 mm*10 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 30%-60%, 10 min). The desired fraction was lyophilizedto yield5-(1,3-benzodioxol-5-yl)-N-[2-(1H-indol-3-yl)ethyl]-3-isopropyl-pyrazolo[1,5-a]pyrimidin-7-amine(22.10 mg, 43.13 μmol, 25.1% yield, 100.0% purity, 2 HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.09 (s, 1H), 7.56 (d, J=7.9 Hz,1H), 7.22 (d, J=8.2 Hz, 1H), 7.08-7.02 (m, 2H), 6.92-6.86 (m, 2H), 6.80(d, J=1.8 Hz, 1H), 6.73 (dd, J=1.9, 8.3 Hz, 1H), 6.08 (s, 2H), 5.70 (s,1H), 3.99-3.93 (m, 2H), 3.28-3.18 (m, 3H), 1.33 (d, J=6.8 Hz, 6H);ES-LCMS m/z 440.2 [M+H]⁺.

Example 189

Synthesis of I-249a, I-249b and I-249c

Synthetic Scheme:

Step 1: 1-(1,4-Dioxaspiro[4.5]dec-7-en-8-yl)pyrrolidine

To a mixture of 1,4-dioxaspiro[4.5]decan-8-one (18 g, 115.25 mmol, 1eq), TsOH (198.47 mg, 1.15 mmol, 0.01 eq) in toluene (200 mL) was addedpyrrolidine (12.30 g, 172.88 mmol, 14.43 mL, 1.5 eq) under N₂atmosphere. The mixture was stirred at 110° C. for 12 h under N₂atmosphere. The reaction mixture was concentrated to yield1-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyrrolidine (24 g, 103.21 mmol,89.6% yield, 90% purity) as brown oil. ¹H NMR (400 MHz, CDCl₃) δ ppm4.17 (t, J=3.7 Hz, 1H), 4.11 (s, 2H), 4.07-4.05 (m, 2H), 3.12 (t, J=6.5Hz, 2H), 2.97 (t, J=6.2 Hz, 2H), 2.59 (t, J=7.1 Hz, 2H), 2.54-2.49 (m,1H), 2.45-2.43 (m, 1H), 2.09 (t, J=7.1 Hz, 2H), 1.94-1.90 (m, 2H),1.87-1.73 (m, 2H).

Step 2: 7-Phenacyl-1,4-dioxaspiro[4.5]decan-8-one

A solution of 1-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)pyrrolidine (24 g,103.21 mmol, 1 eq) in toluene (200 mL) was heated to reflux under N₂atmosphere and 2-bromo-1-phenyl-ethanone (20.54 g, 103.21 mmol, 1 eq) intoluene (60 mL) was added slowly. The mixture was stirred at 110° C. for2 h under N₂ atmosphere. Then H₂O (60 mL) was added and the mixture wasstirred at 110° C. for further 2 h under N₂ atmosphere. The mixture wasconcentrated and water (50 mL) was added, extracted with EtOAc (100mL×3). The combined organic layers were washed with brine (50 mL), driedover Na₂SO₄, filtered and concentrated. The crude material was purifiedon silica gel column chromatography (from pure PE to PE/EtOAc=2/1, TLC:PE/EtOAc=3/1, R_(f)=0.4) to yield7-phenacyl-1,4-dioxaspiro[4.5]decan-8-one (13 g, 42.65 mmol, 41.3%yield, 90% purity) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ ppm8.01-7.93 (m, 2H), 7.59-7.53 (m, 1H), 7.49-7.43 (m, 2H), 4.17-4.06 (m,2H), 4.05-3.96 (m, 2H), 3.64-3.45 (m, 2H), 2.85-2.68 (m, 2H), 2.48-2.39(m, 1H), 2.25-2.14 (m, 1H), 2.13-2.02 (m, 2H), 1.89-1.77 (m, 1H);ES-LCMS m/z 275.1 [M+H]⁺.

Step 3: 2′-Phenylspiro[1,3-dioxolane-2,5′-1,4,6,7-tetrahydroindole]

A mixture of 7-phenacyl-1,4-dioxaspiro[4.5]decan-8-one (5 g, 16.40 mmol,1 eq) and (NH₄)₂CO₃ (5.00 g, 52.04 mmol, 5.56 mL, 3.17 eq) in sealedtube was stirred at 110° C. for 2 h under N₂ atmosphere. To the mixturewas added water (50 mL), extracted with EtOAc (100 mL×2). The combinedorganic layers were washed with brine (40 mL), dried over Na₂SO₄,filtered and concentrated to yield2′-phenylspiro[1,3-dioxolane-2,5′-1,4,6,7-tetrahydroindole] (4 g, 14.10mmol, 86.0% yield, 90% purity) as a brown solid. ¹H NMR (400 MHz, CDCl₃)δ ppm 8.07-7.86 (m, 1H), 7.40 (d, J=7.8 Hz, 2H), 7.36-7.29 (m, 2H),7.20-7.10 (m, 1H), 6.25 (d, J=2.4 Hz, 1H), 4.08-3.99 (m, 4H), 2.84 (t,J=6.6 Hz, 2H), 2.80 (s, 2H), 2.02 (t, J=6.5 Hz, 2H); ES-LCMS m/z 256.1[M+H]⁺.

Step 4: 2-Phenyl-1,4,6,7-tetrahydroindol-5-one

To a solution of2′-phenylspiro[1,3-dioxolane-2,5′-1,4,6,7-tetrahydroindole] (3.9 g,13.75 mmol, 1 eq) in acetone (30 mL) was added HCl (1 M, 13.75 mL, 1eq), the mixture was stirred at 50° C. for 3 h. The mixture wasconcentrated and adjusted pH to 7 with saturated NaHCO₃ solution,extracted with EtOAc (80 mL×2). The combined organic layers were washedwith brine (50 mL), dried over Na₂SO₄, filtered and concentrated. Thecrude material was purified on silica gel column chromatography (frompure PE to PE/EtOAc=2/1, TLC: PE/EtOAc=3/1, R_(f)=0.3) to yield2-phenyl-1,4,6,7-tetrahydroindol-5-one (0.7 g, 2.32 mmol, 16.9% yield,70.0% purity) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.15 (s,1H), 7.44 (d, J=7.6 Hz, 2H), 7.36 (t, J=7.7 Hz, 2H), 7.23-7.17 (m, 1H),6.29 (d, J=2.4 Hz, 1H), 3.45 (s, 2H), 3.09-3.03 (m, 2H), 2.74 (t, J=6.8Hz, 2H); ES-LCMS m/z 212.1 [M+H]⁺.

Step 5: 2-Phenyl-1,4,6,7-tetrahydroindol-5-one oxime

To a solution of 2-phenyl-1,4,6,7-tetrahydroindol-5-one (600 mg, 1.99mmol, 1 eq) in THE (15 mL) was added NaOAc (244.64 mg, 2.98 mmol, 1.5eq) and NH₂OH.HCl (165.78 mg, 2.39 mmol, 1.2 eq). The mixture wasstirred at 60° C. for 3 h. The mixture was filtered. The filtrate wasconcentrated to yield a residue which was purified on silica gel columnchromatography (from pure PE to PE/EtOAc=1/1, TLC: PE/EtOAc=1/1,R_(f)=0.4) to yield 2-phenyl-1,4,6,7-tetrahydroindol-5-one oxime (350mg, 1.24 mmol, 62.2% yield, 80% purity) as a red solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 8.02 (s, 1H), 7.47-7.41 (m, 2H), 7.38-7.33 (m, 2H),7.21-7.16 (m, 1H), 6.32 (d, J=2.4 Hz, 1H), 3.66 (s, 2H), 2.89-2.81 (m,2H), 2.71-2.63 (m, 1H); ES-LCMS m/z 227.1 [M+H]⁺.

Step 6: 2-Phenyl-4,5,6,7-tetrahydro-1H-indol-5-amine

To a solution of 2-phenyl-1,4,6,7-tetrahydroindol-5-one oxime (350 mg,1.24 mmol, 1 eq) in MeOH (6 mL) was added Raney-Ni (200 mg, 2.33 mmol,1.89 eq) under N₂ atmosphere. The mixture was stirred at 20° C. for 1 hunder H₂ (15 psi) atmosphere. The mixture was filtered. The filtrate wasconcentrated to yield 2-phenyl-4,5,6,7-tetrahydro-1H-indol-5-amine (250mg, 942.11 μmol, 76.1% yield, 80% purity) was obtained as a black brownsolid. ¹H NMR (500 MHz, CD₃OD) δ ppm 7.50-7.46 (m, 2H), 7.26 (t, J=7.9Hz, 2H), 7.09-7.03 (m, 1H), 6.18 (s, 1H), 3.12-3.04 (m, 1H), 2.86-2.63(m, 3H), 2.30 (dd, J=8.9, 14.8 Hz, 1H), 2.05-1.97 (m, 1H), 1.73-1.64 (m,1H); ES-LCMS m/z 213.2 [M+H]⁺.

Step 7:2-(5-Fluoro-3-pyridyl)-8-isopropyl-N-[(5R)-2-phenyl-4,5,6,7-tetrahydro-1H-indol-5-yl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-249b) and2-(5-Fluoro-3-pyridyl)-8-isopropyl-N-[(5S)-2-phenyl-4,5,6,7-tetrahydro-1H-indol-5-yl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-249a)

To a solution of 2-phenyl-4,5,6,7-tetrahydro-1H-indol-5-amine (207.84mg, 783.24 umol, 1.2 eq) and DIEA (253.07 mg, 1.96 mmol, 341.07 μL, 3eq) in i-PrOH (15 mL) was added4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(200 mg, 652.70 μmol, 1 eq). The mixture was stirred at 50° C. for 2 h.The mixture was concentrated. The crude material was purified on silicagel column chromatography (from pure PE to PE/EtOAc=2/1, TLC:PE/EtOAc=2/1, R_(f)=0.5) to yield the residue which was separated by SFC(column: DAICEL CHIRALCEL OJ-H (250 mm*30 mm, 5 m); mobile phase: [0.1%NH₃H₂O EtOH]; B %: 40%-40%, min) to yield Peak 1 and Peak 2. Peak 1 wasconcentrated and purified by preparative HPLC (column: Agela ASB 150*25mm*5 m; mobile phase: [water (0.05% HCl)-ACN]; B %: 75%-100%, 8 min),followed by lyophilization to yield an enantiomer (18.68 mg, 31.16 μmol,4.8% yield, 96.249% purity, 3 HCl. SFC: Rt=5.229 min, ee=95.522%,[α]^(26.5) _(D)=+24.339, (CHCl₃, c=0.1015 g/100 mL)) as a yellow solid.¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.89 (s, 1H), 9.38 (s, 1H), 8.89 (d,J=8.6 Hz, 1H), 8.70 (d, J=2.9 Hz, 1H), 8.43 (td, J=2.2, 9.8 Hz, 1H),8.18-8.07 (m, 1H), 7.55 (d, J=7.6 Hz, 2H), 7.30 (t, J=7.7 Hz, 2H),7.14-7.03 (m, 1H), 6.27 (d, J=2.2 Hz, 1H), 4.77-4.60 (m, 1H), 3.26-3.15(m, 1H), 2.93-2.71 (m, 4H), 2.17-2.00 (m, 2H), 1.36 (d, J=7.1 Hz, 6H);ES-LCMS m/z 468.3 [M+H]⁺. Peak 2 was concentrated to yield a residuewhich was dissolved in MeCN (20 mL), 1 M HCl solution (1.2 mL) and H₂O(40 mL), followed by lyophilization to yield the other enantiomer (23.41mg, 40.58 μmol, 6.2% yield, 100% purity, 3 HCl. SFC: Rt=6.074 min,ee=100%, [α]^(26.5) _(D)=−26.002, (CHCl₃, c=0.1002 g/100 mL)) as ayellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.90 (br s, 1H), 9.38 (s,1H), 8.89 (d, J=8.6 Hz, 1H), 8.70 (d, J=2.7 Hz, 1H), 8.43 (d, J=9.8 Hz,1H), 8.23-8.04 (m, 1H), 7.55 (d, J=7.6 Hz, 2H), 7.30 (t, J=7.8 Hz, 2H),7.17-7.03 (m, 1H), 6.27 (d, J=2.2 Hz, 1H), 4.68 (d, J=5.1 Hz, 1H),3.27-3.13 (m, 1H), 2.92-2.66 (m, 4H), 2.19-2.00 (m, 2H), 1.36 (d, J=6.8Hz, 6H); ES-LCMS m/z 468.3 [M+H]⁺.

Example 190

Synthesis of I-252a, I-252b and I-252c

Synthetic Scheme:

Step 1:2-Chloro-N-(4,5,6,7-tetrahydro-1H-indol-5-yl)pyrido[3,2-d]pyrimidin-4-amine

A mixture of 2,4-dichloropyrido[3,2-d]pyrimidine (150 mg, 749.91 μmol, 1eq), 4,5,6,7-tetrahydro-1H-indol-5-amine (113.48 mg, 749.91 mol, 1 eq)and DIEA (387.68 mg, 3.00 mmol, 522.48 μL, 4 eq) in i-PrOH (10 mL) wasstirred at 55° C. for 2 h. The reaction mixture was concentrated toyield the residue which was purified on silica gel column chromatography(from PE/EtOAc=3/1 to 1/1, TLC: PE/EtOAc=1/1, R_(f)=0.50) to yield2-chloro-N-(4,5,6,7-tetrahydro-1H-indol-5-yl)pyrido[3,2-d]pyrimidin-4-amine(120 mg, 397.52 μmol, 53.0% yield, 99.3% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.64 (dd, J=1.5, 4.3 Hz, 1H), 8.01 (dd,J=1.5, 8.5 Hz, 1H), 7.85 (s, 1H), 7.64 (dd, J=4.3, 8.3 Hz, 1H), 7.44 (d,J=8.5 Hz, 1H), 6.70 (t, J=2.6 Hz, 1H), 6.04 (t, J=2.6 Hz, 1H), 4.80-4.70(m, 1H), 3.10 (dd, J=5.3, 15.3 Hz, 1H), 2.99-2.45 (m, 3H), 2.24-2.08 (m,2H); ES-LCMS m/z 300.1 [M+H]⁺.

Step 2:2-(5-Fluoro-3-pyridyl)-N-[(5R)-4,5,6,7-tetrahydro-1H-indol-5-yl]pyrido[3,2-d]pyrimidin-4-amine(I-252b) and2-(5-Fluoro-3-pyridyl)-N-[(5S)-4,5,6,7-tetrahydro-1H-indol-5-yl]pyrido[3,2-d]pyrimidin-4-amine(I-252a)

2-Chloro-N-(4,5,6,7-tetrahydro-1H-indol-5-yl)pyrido[3,2-d]pyrimidin-4-amine(120 mg, 397.52 μmol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (168.04mg, 1.19 mmol, 3 eq), Pd(dppf)Cl₂ (29.09 mg, 39.75 μmol, 0.1 eq) andCs₂CO₃ (518.08 mg, 1.59 mmol, 4 eq) in 1,4-dioxane (3 mL) and H₂O (1 mL)were taken up into a microwave tube and purged with N₂ atmosphere for 1min. The sealed tube was heated at 110° C. for 1 h under microwave. Thereaction mixture was concentrated. The residue was purified by flashsilica gel chromatography (from pure PE to PE/EtOAc=1/1, TLC:PE/EtOAc=2/1, R_(f)=0.40) to yield a residue which was separated bychiral SFC (column: DAICEL CHIRALCEL OJ-H (250 mm*30 mm, 5 μm); mobilephase: [0.1% NH₃H₂O EtOH]; B %: 40%-40%) to yield an enantiomer (19.36mg, 52.04 μmol, 13.1% yield, 96.9% purity, SFC: Rt=4.040, ee=100%,Optical rotation: [α]^(23.9) _(D)=−8.542 (CHCl₃, c=0.075 g/100 mL)) as ared solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.43 (s, 1H), 8.71 (dd, J=1.5,4.2 Hz, 1H), 8.56-8.48 (m, 2H), 8.15 (dd, J=1.5, 8.6 Hz, 1H), 7.76-7.73(m, 1H), 6.62 (d, J=2.4 Hz, 1H), 5.90 (d, J=2.4 Hz, 1H), 4.84-4.78 (m,1H), 3.09 (dd, J=4.9, 15.2 Hz, 1H), 2.85-2.69 (m, 3H), 2.27 (d, J=12.5Hz, 1H), 2.14 (d, J=6.6 Hz, 1H); ES-LCMS m/z 361.2 [M+H]⁺ and the otherenantiomer (17.54 mg, 47.86 μmol, 12.0% yield, 98.3% purity) as a redsolid (SFC: Rt=4.941, ee=100%, Optical rotation: [α]^(25.0) _(D)=+10.973(CHCl₃, c=0.054 g/100 mL)). ¹H NMR (400 MHz, CD₃OD) δ ppm 9.44 (s, 1H),8.71 (dd, J=1.2, 4.2 Hz, 1H), 8.57-8.49 (m, 2H), 8.16 (dd, J=1.2, 8.6Hz, 1H), 7.75 (dd, J=4.2, 8.6 Hz, 1H), 6.64 (s, 1H), 5.92 (d, J=2.4 Hz,1H), 4.86-4.81 (m, 1H), 3.11 (dd, J=5.0, 15.0 Hz, 1H), 2.88-2.70 (m,3H), 2.28 (d, J=9.8 Hz, 1H), 2.19-2.11 (m, 1H); ES-LCMS m/z 361.2[M+H]⁺.

Example 191

Synthesis of I-253

Synthetic Scheme:

Step 1:(3R)—N-[2-(5-Fluoro-3-pyridyl)-8-(2-tetrahydropyran-2-ylpyrazol-3-yl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine

(3R)—N-[8-bromo-2-(5-fluoro-3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(100 mg, 202.79 μmol, 1 eq), Cs₂CO₃ (66.07 mg, 202.79 μmol, 1 eq),Pd(dppf)Cl₂ (148.39 mg, 202.79 μmol, 1 eq) and1-tetrahydropyran-2-yl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole(56.41 mg, 202.79 μmol, 1 eq) were taken up into a microwave tube in1,4-dioxane (6 mL) and H₂O (2 mL). The sealed tube was heated at 110° C.for 30 min under microwave. TLC (PE/EtOAc=1/1, R_(f)=0.4) indicated thestarting material was consumed completely. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by flash silica gel chromatography (from PE/EA=1/0 to 1/1, TLC:PE/EtOAc=1/1, R_(f)=0.4) to yield(3R)—N-[2-(5-fluoro-3-pyridyl)-8-(2-tetrahydropyran-2-ylpyrazol-3-yl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(70 mg, 122.27 μmol, 60.3% yield, 96.0% purity) as a yellow solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 10.82 (s, 1H), 9.41-9.38 (m, 2H), 8.73 (d,J=2.7 Hz, 1H), 8.49-8.47 (d, J=9.0 Hz, 1H), 8.38 (s, 1H), 7.64 (s, 1H),7.37-7.35 (d, J=7.8 Hz, 1H), 7.29-7.27 (d, J=7.8 Hz, 1H), 7.02-6.99 (m,1H), 6.95-6.92 (m, 1H), 6.74 (s, 1H), 5.66-5.64 (d, J=9.0 Hz, 1H), 4.89(d, 1H), 3.95-3.91 (m, 2H), 3.59-3.57 (m, 1H), 3.15-2.88 (m, 4H), 2.23(br s, 2H), 1.64-1.52 (m, 3H), 1.23 (m, 1H), 0.85-0.83 (d, J=7.8 Hz,1H); ES-LCMS m/z 550.3 [M+H]⁺.

Step 2:(3R)—N-[2-(5-Fluoro-3-pyridyl)-8-(1H-pyrazol-5-yl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine (I-253)

A solution of(3R)—N-[2-(5-fluoro-3-pyridyl)-8-(2-tetrahydropyran-2-ylpyrazol-3-yl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine (70mg, 122.27 μmol, 1 eq) in HCl/MeOH (10 mL) was stirred at 15° C. for 20min. The reaction mixture was basified with aqueous NaHCO₃ solutionuntil pH to 7-8, extracted with EtOAc (50 mL×3), dried over Na₂SO₄,filtered and concentrated under reduced pressure to yield a residuewhich was purified by preparative HPLC (HCl condition; column: Agela ASB150*25 mm*5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 53%-83%, 8min) to yield(3R)—N-[2-(5-fluoro-3-pyridyl)-8-(1H-pyrazol-5-yl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(19.45 mg, 33.83 μmol, 27.7% yield, 100.0% purity, 3HCl, [α]^(26.4)_(D)=+13.329 (MeOH, c=0.098 g/100 mL)) as a yellow solid. ¹H NMR (400MHz, DMSO-d₆) δ ppm 10.82 (s, 1H), 9.51 (s, 1H), 9.29-9.27 (d, J=8.6 Hz,1H), 8.75 (d, J=2.7 Hz, 1H), 8.68 (d, J=10.0 Hz, 1H), 8.64 (s, 1H),7.84-7.83 (d, J=2.0 Hz, 1H), 7.37-7.35 (d, J=7.6 Hz, 1H), 7.29-7.27 (d,J=8.1 Hz, 1H), 7.04-7.00 (m, 2H), 6.95-6.93 (m, 1H), 4.89-4.87 (d, J=7.6Hz, 1H), 3.17-3.12 (m, J=5.5, 14.8 Hz, 1H), 3.04-2.88 (m, 3H), 2.24-2.23(d, J=3.7 Hz, 2H); ES-LCMS m/z 466.2 [M+H]⁺.

Example 192

Synthesis of I-254

Synthetic Scheme:

Step 1:tert-Butyl(3R)-3-tert-butoxycarbonyl-(8-isopropyl-2-morpholino-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino-1,2,3,4-tetrahydrocarbazole-9-carboxylate

tert-Butyl-(3R)-3-[tert-butoxycarbonyl-(8-isopropyl-2-methylsulfinyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(100 mg, 156.06 μmol, 1 eq) and morpholine (40.79 mg, 468.17 μmol, 41.20μL, 3 eq) in t-BuOH (4 mL) were taken up into a microwave tube andpurged with N₂ for 1 min. The sealed tube was heated at 120° C. for 2 hunder microwave. The reaction mixture was concentrated to yieldtert-butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropyl-2-morpholino-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(100 mg, 142.46 μmol, 91.3% yield, 90% purity) as yellow oil which wasused in the next step without purification. ¹H NMR (400 MHz, CDCl₃) δppm 8.10 (d, J=8.0 Hz, 1H), 7.82 (s, 1H), 7.35 (d, J=7.5 Hz, 1H), 7.22(dd, J=6.7, 13.9 Hz, 2H), 4.74 (s, 1H), 3.79-3.74 (m, 8H), 3.23 (d,J=10.8 Hz, 2H), 3.15-3.07 (m, 3H), 2.26 (d, J=4.3 Hz, 2H), 1.65 (s, 9H),1.36 (s, 9H), 1.33 (d, J=7.0 Hz, 6H); ES-LCMS m/z 632.3 [M+H]⁺.

Step 2:(3R)—N-(8-Isopropyl-2-morpholino-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-254)

To a solution of tert-butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropyl-2-morpholino-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(100 mg, 142.46 μmol, 1 eq) in DCM (4 mL) was added TFA (1.54 g, 13.51mmol, 1 mL, 94.81 eq). The reaction mixture was stirred at 20° C. for0.5 h. The reaction mixture was concentrated. The residue was purifiedby preparative HPLC (column: Agela ASB 150*25 mm*5 μm; mobile phase:[water (0.05% HCl)-ACN]; B %: 50%-80%, 8 min), followed bylyophilization to yield(3R)—N-(8-isopropyl-2-morpholino-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(28.19 mg, 59.32 μmol, 41.6% yield, 98.5% purity, HCl, [α]^(24.7)_(D)=+84.327, MeOH, c=0.074 g/100 mL) as a white solid. ¹H NMR (400 MHz,CD₃OD) δ ppm 7.95 (s, 1H), 7.37 (d, J=7.8 Hz, 1H), 7.28 (d, J=8.0 Hz,1H), 7.08-7.02 (m, 1H), 7.00-6.94 (m, 1H), 4.72-4.63 (m, 1H), 3.93-3.87(m, 4H), 3.83-3.78 (m, 4H), 3.27-3.19 (m, 2H), 3.03-2.86 (m, 3H),2.35-2.18 (m, 2H), 1.32 (d, J=6.8 Hz, 6H); ES-LCMS m/z 432.3 [M+H]⁺.

Example 193

Synthesis of I-255

Synthetic Scheme:

Step 1:(3R)—N-[8-Isopropyl-2-(3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-255)

(3R)—N-(8-isopropyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(100 mg, 242.03 μmol, 1 eq), 3-pyridylboronic acid (89.25 mg, 726.08μmol, 3 eq), Pd(PPh₃)₄ (27.97 mg, 24.20 μmol, 0.1 eq) andthiophene-2-carbonyloxycopper (138.46 mg, 726.08 μmol, 3 eq) in1,4-dioxane (3 mL) were taken up into a microwave tube and purged withN₂ for 1 min. The sealed tube was heated at 120° C. for 2 h undermicrowave. The reaction mixture was filtered and concentrated to givethe residue which was purified by preparative HPLC (column: Xtimate C18150*25 mm*5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 35%-65%, 8min) twice, followed by lyophilization to yield(3R)—N-[8-isopropyl-2-(3-pyridyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(28.39 mg, 56.08 μmol, 23.2% yield, 98.1% purity, 2 HCl, [α]²⁵_(D)=+34.261 (MeOH, c=0.071 g/100 mL)) as a yellow solid. ¹H NMR (400MHz, CD₃OD) δ ppm 9.69 (s, 1H), 9.49 (d, J=8.3 Hz, 1H), 8.89 (d, J=4.5Hz, 1H), 8.16-8.10 (m, 1H), 8.06 (s, 1H), 7.38 (d, J=7.8 Hz, 1H), 7.29(d, J=8.0 Hz, 1H), 7.08-7.03 (m, 1H), 7.00-6.94 (m, 1H), 4.90-4.88 (m,1H), 3.37-3.34 (m, 1H), 3.30 (d, J=7.0 Hz, 1H), 3.13-2.90 (m, 3H),2.44-2.24 (m, 2H), 1.45 (d, J=7.0 Hz, 6H); ES-LCMS m/z 424.3 [M+H]⁺.

Example 194

Synthesis of I-256

Synthetic Scheme:

Step 1: Ethyl 1-aminoimidazole-2-carboxylate

To a solution of ethyl 1H-imidazole-2-carboxylate (1 g, 7.14 mmol, 1 eq)in DMF (80 mL) cooled to −10° C. was added LiHMDS (1 M, 7.85 mL, 1.1eq). The mixture was stirred at −10° C. for 0.5 h. To the mixture wasadded N-diphenylphosphorylhydroxylamine (1.83 g, 7.85 mmol, 1.1 eq). Themixture was stirred at 15° C. for 11.5 h. The mixture was quenched withsaturated aqueous NH₄Cl solution (50 mL) at −10° C. The mixture wasconcentrated and to the residue was added EtOAc (100 mL). Afterfiltration, washed with EtOAc (50 mL×3), the filtrate was concentratedto yield ethyl 1-aminoimidazole-2-carboxylate (3 g, crude) as a yellowsolid which was used in the next step without further purification. ¹HNMR (400 MHz, CDCl₃) δ ppm 7.10 (s, 1H), 6.98 (s, 1H), 5.74 (s, 2H),4.33 (q, J=7.2 Hz, 2H), 1.36-1.33 (m, 3H); ES-LCMS m/z 156.0 [M+H]⁺.

Step 2: Ethyl 1-[bis(ethoxycarbonyl)amino]imidazole-2-carboxylate

To a solution of ethyl 1-aminoimidazole-2-carboxylate (5.3 g, 17.08mmol, 1 eq) in THE (80 mL) and H₂O (80 mL) was added Na₂CO₃ (12.67 g,119.56 mmol, 7 eq) and ethyl carbonochloridate (8.47 g, 78.05 mmol, 7.43mL, 4.57 eq). The mixture was stirred at 15° C. for 2 h. To the mixturewas added H₂O (100 mL), extracted with EtOAc (100 mL×2). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated. Theresidue was purified on silica gel column chromatography (from pure PEto PE/EtOAc=3/1, TLC: PE/EtOAc=3/1, R_(f)=0.6) to yield ethyl1-[bis(ethoxycarbonyl)amino]imidazole-2-carboxylate (2.69 g, 6.29 mmol,36.8% yield, 70% purity) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δppm 7.30 (d, J=1.0 Hz, 1H), 7.10 (s, 1H), 4.44-4.33 (m, 4H), 4.19-4.08(m, 2H), 1.39 (td, J=7.3, 16.3 Hz, 6H), 1.08 (t, J=7.2 Hz, 3H); ES-LCMSm/z 300.1 [M+H]⁺.

Step 3: 1H-Imidazo[2,1-f][1,2,4]triazine-2,4-dione

A solution of ethyl 1-[bis(ethoxycarbonyl)amino]imidazole-2-carboxylate(2.1 g, 4.91 mmol, 1 eq) and NH₃.H₂O (97.50 g, 778.99 mmol, 107.14 mL,158.59 eq) in i-PrOH (20 mL) was stirred in a sealed tube at 120° C. for16 h. The mixture was concentrated. To the residue was added PE/MeOH(10/1, 50 mL) and stirred at room temperature for 10 min. Afterfiltration, washed with PE/MeOH (10/1, 15 mL), the filter cake was driedin vacuo. The crude product (1 g) was purified on silica gel columnchromatography (from DCM/MeOH=10/1 to 1/2, TLC: DCM/MeOH=10/1,R_(f)=0.3) to yield 1H-imidazo[2,1-f][1,2,4]triazine-2,4-dione (0.9 g,96.4% yield, 80.0% purity). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.02 (br s,1H), 7.24 (s, 1H), 7.13 (br s, 1H), 7.07 (s, 1H).

Step 4: 2,4-Dichloroimidazo[2,1-f][1,2,4]triazine

A mixture of 1H-imidazo[2,1-f][1,2,4]triazine-2,4-dione (500 mg, 2.63mmol, 1 eq), N,N-diethylethanamine, hydrochloride (723.95 mg, 5.26 mmol,2 eq) and POCl₃ (87.18 g, 568.57 mmol, 52.84 mL, 216.21 eq) was stirredin a sealed tube at 120° C. for 24 h. The mixture was concentrated andto the residue was added DCM (30 mL). The mixture was poured into icecold water (50 mL), extracted with DCM (30 mL×2). The combined organiclayers were washed with brine (50 mL), dried over Na₂SO₄, filtered andconcentrated to yield 2,4-dichloroimidazo[2,1-f][1,2,4]triazine (400 mg,1.27 mmol, 48.3% yield, 60% purity) as a yellow solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 8.09 (s, 1H), 8.02 (s, 1H); ES-LCMS m/z 189.2, 191.1[M+H]⁺.

Step 5:(3R)—N-(2-chloroimidazo[2,1-f][1,2,4]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine

A mixture of 2,4-dichloroimidazo[2,1-f][1,2,4]triazine (400 mg, 1.27mmol, 1 eq), DIEA (492.34 mg, 3.81 mmol, 663.53 μL, 3 eq) and(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (236.51 mg, 1.27 mmol, 1 eq)in CH₃CN (15 mL) was stirred at 70° C. for 6 h. The mixture wasconcentrated. The residue was purified on silica gel columnchromatography (from PE/EtOAc=5/1 to 1/2, TLC: PE/EtOAc=1/1, R_(f)=0.4)to yield(3R)—N-(2-chloroimidazo[2,1-f][1,2,4]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(220 mg, 636.38 μmol, 50.1% yield, 98.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 7.83 (br s, 1H), 7.67 (s, 1H), 7.48-7.36 (m,2H), 7.30 (d, J=7.8 Hz, 1H), 7.18-7.12 (m, 1H), 7.11-7.04 (m, 1H), 6.84(d, J=7.3 Hz, 1H), 4.86-4.84 (m, 1H), 3.27 (dd, J=4.9, 15.7 Hz, 1H),2.98-2.80 (m, 3H), 2.29-2.17 (m, 2H); ES-LCMS m/z 339.1 [M+H]⁺.

Step 6:(3R)—N-[2-(5-fluoro-3-pyridyl)imidazo[2,1-f][1,2,4]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-256)

A mixture of(3R)—N-(2-chloroimidazo[2,1-f][1,2,4]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(100 mg, 289.26 μmol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (81.52 mg,578.52 μmol, 2 eq), Cs₂CO₃ (282.74 mg, 867.78 μmol, 3 eq) andPd(dppf)Cl₂ (21.17 mg, 28.93 μmol, 0.1 eq) in 1,4-dioxane (2 mL) and H₂O(0.5 mL) was stirred at 110° C. for 2 h under N₂ atmosphere. Afterfiltration, the filtrate was concentrated. The residue was purified bypreparative TLC (PE/EtOAc=1/1, R_(f)=0.3) to yield(3R)—N-[2-(5-fluoro-3-pyridyl)imidazo[2,1-f][1,2,4]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(60.97 mg, 152.65 μmol, 52.8% yield, 100% purity, [α]^(26.9) _(D)=+9.272(MeOH, c=0.101 g/100 mL)) as a yellow solid. ¹H NMR (400 MHz, CD₃OD) δppm 9.30 (s, 1H), 8.54 (d, J=2.7 Hz, 1H), 8.42 (td, J=2.1, 9.8 Hz, 1H),7.96 (d, J=1.0 Hz, 1H), 7.55 (s, 1H), 7.36 (d, J=7.6 Hz, 1H), 7.25 (d,J=8.1 Hz, 1H), 7.05-6.97 (m, 1H), 6.96-6.89 (m, 1H), 4.98-4.90 (m, 1H),3.26 (br s, 1H), 2.97 (t, J=5.7 Hz, 2H), 2.90 (dd, J=7.2, 15.3 Hz, 1H),2.40-2.32 (m, 1H), 2.31-2.21 (m, 1H); ES-LCMS m/z 399.9 [M+H]⁺.

Example 195

Synthesis of I-257

Synthetic Scheme:

Step 1:2-Chloro-N-[2-(1H-indol-3-yl)ethyl]pyrido[3,2-d]pyrimidin-4-amine

To solution of 2,4-di chloropyrido[3,2-d]pyrimidine (100 mg, 499.94μmol, 1 eq) in THE (3 mL) was added DIEA (193.84 mg, 1.50 mmol, 261.24μL, 3 eq) and 2-(1H-indol-3-yl)ethanamine (88.11 mg, 549.93 μmol, 1.1eq). The mixture was stirred at 60° C. for 2 h. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified by flash silica gel chromatography (from PE/EtOAc=100/1 to 3/1,TLC: PE/EtOAc=3/1, R_(f)=0.2) to yield2-chloro-N-[2-(1H-indol-3-yl)ethyl]pyrido[3,2-d]pyrimidin-4-amine (90mg, 271.57 μmol, 54.30 yield, 97.7% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.61 (dd, J=1.3, 4.3 Hz, 1H), 8.07 (s, 1H), 8.01(dd, J=1.3, 8.4 Hz, 1H), 7.71 (d, J=7.8 Hz, 1H), 7.63 (dd, J=4.3, 8.4Hz, 1H), 7.47 (s, 1H), 7.41 (d, J=8.1 Hz, 1H), 7.24 (t, J=7.5 Hz, 1H),7.18-7.12 (m, 2H), 4.03 (q, J=6.7 Hz, 2H), 3.22 (t, J=6.8 Hz, 2H);ES-LCMS m/z 324.1 [M+H]⁺.

Step 2:2-(5-Fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrido[3,2-d]pyrimidin-4-amine(I-257)

2-Chloro-N-[2-(1H-indol-3-yl)ethyl]pyrido[3,2-d]pyrimidin-4-amine (50mg, 150.87 μmol, 1 eq), (5-fluoro-3-pyridyl)boronic acid (25.51 mg,181.05 μmol, 1.2 eq), Pd(dppf)Cl₂ (11.04 mg, 15.09 μmol, 0.1 eq) andCs₂CO₃ (147.47 mg, 452.62 μmol, 3 eq) were taken up into a microwavetube in 1,4-dioxane (2 mL) and H₂O (1 mL). The sealed tube was heated at110° C. for 0.5 h under microwave. The reaction mixture was diluted withwater (5 mL) and extracted with EtOAc (10 mL×3). The combined organiclayers were dried over Na₂SO₄, filtered and concentrated under reducedpressure to yield a residue which was purified by preparative HPLC(column: Xtimate C18 150*25 mm*5 μm; mobile phase: [water (0.05%HCl)-ACN]; B %: 45%-75%, 8 min), followed by lyophilization to yield2-(5-fluoro-3-pyridyl)-N-[2-(1H-indol-3-yl)ethyl]pyrido[3,2-d]pyrimidin-4-amine(24.02 mg, 51.42 μmol, 34.1% yield, 97.9% purity, 2HCl) as a yellowsolid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.03 (s, 1H), 9.01 (dd, J=1.3, 4.3Hz, 1H), 8.82 (d, J=2.7 Hz, 1H), 8.20 (dd, J=1.5, 8.6 Hz, 1H), 8.10-8.05(m, 1H), 8.01 (dd, J=4.3, 8.7 Hz, 1H), 7.56 (d, J=7.8 Hz, 1H), 7.14 (d,J=8.1 Hz, 1H), 7.05 (s, 1H), 6.92 (t, J=7.2 Hz, 1H), 6.85-6.71 (m, 1H),4.26 (t, J=6.6 Hz, 2H), 3.26 (t, J=6.6 Hz, 2H); ES-LCMS m/z 385.2[M+H]⁺.

Example 196

Synthesis of I-258

Synthetic Scheme:

Step 1:(3R)—N-[2-(3,6-Dihydro-2H-pyran-4-yl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine

To a stirred solution of(3R)—N-(8-isopropyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(300 mg, 703.15 μmol, 1 eq) and2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(443.15 mg, 2.11 mmol, 3 eq) in 1,4-dioxane (15 mL) was degassed andpurged with N₂ then added thiophene-2-carbonyloxycopper (268.17 mg, 1.41mmol, 2 eq) and Pd₂(dba)₃ (64.39 mg, 70.31 μmol, 0.1 eq). The reactionmixture was stirred at 130° C. for 12 h under N₂ atmosphere. Thereaction mixture was filtered through a pad of celite. The filtrate wasconcentrated to yield a residue which was purified by flash silica gelchromatography (from PE/EtOAc=100/1 to 3/1, TLC: PE/EtOAc=3/1,R_(f)=0.55) to yield(3R)—N-[2-(3,6-dihydro-2H-pyran-4-yl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(60 mg, 127.97 μmol, 18.2% yield, 91.4% purity) as a white solid. ¹H NMR(400 MHz, CD₃OD) δ ppm 7.87 (s, 1H), 7.37 (d, J=7.6 Hz, 1H), 7.29-7.20(m, 2H), 7.03 (t, J=7.1 Hz, 1H), 6.98-6.92 (m, 1H), 4.73 (s, 1H), 4.34(d, J=2.7 Hz, 2H), 3.89 (t, J=5.5 Hz, 2H), 3.29-3.15 (m, 2H), 3.00-2.91(m, 2H), 2.86 (dd, J=8.9, 15.0 Hz, 1H), 2.69 (d, J=1.7 Hz, 2H), 2.32 (s,1H), 2.26-2.17 (m, 1H), 1.35 (d, J=7.1 Hz, 6H); ES-LCMS m/z 429.3[M+H]⁺.

Step 2:(3R)—N-(8-Isopropyl-2-tetrahydropyran-4-yl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-258)

To a stirred solution of(3R)—N-[2-(3,6-dihydro-2H-pyran-4-yl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(55 mg, 117.31 μmol, 1 eq) in MeOH (20 mL) was added Pd/C (0.1 g, 10%wt). The reaction mixture was stirred at 20° C. for 5 h under H₂atmosphere (15 psi). The reaction mixture was filtered through a pad ofcelite. The filtrate was concentrated to yield a residue which waspurified by preparative HPLC (HCl condition; column: Agela ASB 150*25mm*5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %: 55%-85%, 8 min).The desired fraction was lyophilized to yield(3R)—N-(8-isopropyl-2-tetrahydropyran-4-yl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(16.60 mg, 36.74 μmol, 31.3% yield, 95.3% purity, [α]^(25.0) _(D)=+9.739(MeOH, c=0.098 g/100 mL)) as a white solid. ¹H NMR (400 MHz, CD₃OD) δppm 8.08 (s, 1H), 7.37 (d, J=7.8 Hz, 1H), 7.27 (d, J=8.0 Hz, 1H),7.07-7.01 (m, 1H), 6.99-6.93 (m, 1H), 4.86-4.77 (m, 1H), 4.05 (dd,J=2.0, 11.5 Hz, 2H), 3.61-3.50 (m, 2H), 3.28-3.17 (m, 2H), 3.10-2.88 (m,4H), 2.39-2.20 (m, 2H), 2.11-1.98 (m, 2H), 1.96-1.87 (m, 2H), 1.35 (d,J=6.8 Hz, 6H); ES-LCMS m/z 431.3 [M+H]⁺.

Example 197

Synthesis of I-259a, I-259b and I-259c

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-8-methyl-N-[(5R)-4,5,6,7-tetrahydro-1H-indol-5-yl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-259b) and2-(5-fluoro-3-pyridyl)-8-methyl-N-[(5S)-4,5,6,7-tetrahydro-1H-indol-5-yl]pyrazolo[1,5-a][1,3,5]triazin-4-amine(I-259a)

A mixture of4-chloro-2-(5-fluoro-3-pyridyl)-8-methyl-pyrazolo[1,5-a][1,3,5]triazine(100 mg, 379.28 μmol, 1 eq), 4,5,6,7-tetrahydro-1H-indol-5-amine (56.82mg, 417.21 μmol, 1.1 eq) and DIEA (147.06 mg, 1.14 mmol, 198.19 μL, 3eq) in i-PrOH (3 mL) was stirred at 60° C. for 2 h. The reaction mixturewas concentrated under reduced pressure to yield a residue which waspurified by flash silica gel chromatography (from PE/EtOAc=1/0 to 2/1,TLC: PE/EtOAc=1/1, R_(f)=0.60) to yield crude product which wasseparated by chiral SFC (column: DAICEL CHIRALPAK AD (250 mm*50 mm, 10um); mobile phase: [0.1% NH₃H₂O/IPA]; B %: 40%-40%) to yield Peak 1 andPeak 2. Peak 1 was concentrated under reduced pressure to yield aresidue which was dissolved in MeCN (20 mL) and H₂O (40 mL) andlyophilized to yield an enantiomer (20.50 mg, 56.41 μmol, 14.9% yield,100.0% purity, SFC: R_(t)=1.488, ee=100%, [α]^(26.1) _(D)=−32.731(CHCl₃, c=0.108 g/100 mL)) as an off-white solid. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.51 (s, 1H), 8.56 (d, J=2.8 Hz, 1H), 8.50-8.41 (m, 1H),7.94-7.87 (m, 1H), 7.85 (s, 1H), 6.72 (t, J=2.4 Hz, 1H), 6.66 (d, J=8.0Hz, 1H), 6.05 (t, J=2.4 Hz, 1H), 4.88-4.77 (m, 1H), 3.17 (dd, J=4.8,15.2 Hz, 1H), 2.88-2.71 (m, 3H), 2.35 (s, 3H), 2.28 (dq, J=2.4, 6.0 Hz,1H), 2.23-2.12 (m, 1H); ES-LCMS m/z 364.2 [M+H]⁺. Peak 2 wasconcentrated under reduced pressure to yield a residue which wasdissolved in MeCN (20 mL) and H₂O (40 mL) and lyophilized to yield theother enantiomer (19.32 mg, 53.17 μmol, 14.0% yield, 100.0% purity, SFC:R_(t)=1.691, ee=100%, [α]^(26.0) _(D)=+10.595 (CHCl₃, c=0.102 g/100 mL))as an off-white solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 9.51 (br s, 1H),8.56 (br s, 1H), 8.46 (d, J=9.2 Hz, 1H), 7.90 (s, 1H), 7.85 (s, 1H),6.72 (s, 1H), 6.66 (d, J=8.4 Hz, 1H), 6.05 (s, 1H), 4.90-4.80 (m, 1H),3.16 (d, J=10.8 Hz, 1H), 2.87-2.73 (m, 3H), 2.35 (s, 3H), 2.30-2.22 (m,1H), 2.20-2.10 (m, 1H); ES-LCMS m/z 364.2 [M+H]⁺.

Example 198

Synthesis of I-262

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carboxylate

A solution of2-(5-fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carbonitrile(200 mg, 335.51 μmol, 1 eq) in HCl/MeOH (4 M, 10 mL) was stirred at 70°C. for 12 h in sealed tube. The reaction mixture was concentrated underreduced pressure to give a residue which was quenched by addition ofwater (50 mL), adjusted pH to 9 by aqueous NaHCO₃, extracted with EtOAc(50 mL×3). The combined organic layers were washed with brine (20 mL),dried over Na₂SO₄, filtered and concentrated under reduced pressure toyield a residue which was purified by preparative TLC (PE/EtOAc=1/2,TLC: PE/EtOAc=1/2, R_(f)=0.53) to yield methyl2-(5-fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carboxylate(50 mg, 96.84 μmol, 28.8% yield, 88.6% purity) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ ppm 9.57 (br s, 1H), 8.67-8.48 (m, 2H), 8.41 (s, 1H),7.98 (s, 1H), 7.55-7.42 (m, 1H), 7.36 (d, J=8.1 Hz, 1H), 7.19 (t, J 7.7Hz, 1H), 7.15-7.10 (m, 1H), 6.96 (d, J=8.1 Hz, 1H), 4.99 (s, 1H), 3.98(s, 3H), 3.39 (dd, J=4.8, 15.3 Hz, 1H), 3.09-2.97 (m, 3H), 2.43-2.32 (m,2H); ES-LCMS m/z 458.2 [M+H]⁺.

Step 2:2-(5-Fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carboxylicacid

To a solution of methyl2-(5-fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carboxylate(140 mg, 306.04 μmol, 1 eq) in THE (2 mL), MeOH (2 mL) and H₂O (1 mL)was added LiOH (73.29 mg, 3.06 mmol, 10 eq) was stirred at 50° C. for 12h. The solution was adjusted pH to 6 by 2N aqueous HCl solution,concentrated to yield2-(5-fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carboxylicacid (120 mg, 187.00 μmol, 61.1% yield, 69.1% purity) as a yellow solidwhich was used in the next step without further purification. ¹H NMR(400 MHz, CD₃OD) δ ppm 9.69 (s, 1H), 9.21 (d, J=6.6 Hz, 1H), 8.93 (s,1H), 8.49 (s, 1H), 7.36 (d, J=7.8 Hz, 1H), 7.29-7.17 (m, 1H), 7.08-6.89(m, 2H), 4.93 (d, J=4.2 Hz, 1H), 3.30-3.20 (m, 1H), 3.19-2.88 (m, 3H),2.53-2.08 (m, 2H); ES-LCMS m/z 444.1 [M+H]⁺.

Step 3:[2-(5-Fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazin-8-yl]-morpholino-methanone(I-262)

To a solution of2-(5-fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carboxylicacid (50 mg, 77.91 μmol, 1 eq), morpholine (20.36 mg, 233.74 μmol, 20.57μL, 3 eq) in DMF (2 mL) was added HATU (35.55 mg, 93.50 μmol, 1.2 eq)and TEA (23.65 mg, 233.74 μmol, 32.53 μL, 3 eq). The mixture was stirredat 15° C. for 12 h. The reaction mixture was quenched by addition ofwater (50 mL), extracted with EtOAc (30 mL×3). The combined organiclayers were washed with brine (10 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to yield a residue which waspurified by preparative HPLC (column: DuraShell 150*25 mm*5 μm; mobilephase: [water (0.05% HCl)-ACN]; B %: 32%-62%, 7 min), followed bylyophilization to yield[2-(5-fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazin-8-yl]-morpholino-methanone(15.18 mg, 25.93 μmol, 33.2% yield, 100% purity, 2HCl, [α]^(24.3)_(D)=+20.863 (MeOH, c=0.059 g/100 mL)) as a yellow solid. ¹H NMR (400MHz, CD₃OD) δ ppm 9.52 (s, 1H), 8.90 (dd, J=1.5, 9.0 Hz, 1H), 8.85 (s,1H), 8.35 (s, 1H), 7.37 (d, J=7.6 Hz, 1H), 7.28 (d, J=8.1 Hz, 1H), 7.04(t, J=7.0 Hz, 1H), 6.99-6.93 (m, 1H), 4.99-4.92 (m, 1H), 3.81 (s, 8H),3.29 (d, J=5.6 Hz, 1H), 3.17-2.91 (m, 3H), 2.46-2.23 (m, 2H); ES-LCMSm/z 513.3 [M+H]⁺.

Example 199

Synthesis of I-263a, I-263b and I-263c

Synthetic Scheme:

Step 1:(2S)—N-[2-(5-Fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-1,2,3,4-tetrahydrocyclopenta[b]indol-2-amine(I-263a) and(2R)—N-[2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-1,2,3,4-tetrahydrocyclopenta[b]indol-2-amine(I-263b)

To a solution of4-chloro-2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazine(120 mg, 391.62 μmol, 1 eq) and1,2,3,4-tetrahydrocyclopenta[b]indol-2-amine (67.45 mg, 391.62 μmol, 1eq) in i-PrOH (10 mL) was added DIEA (404.91 mg, 3.13 mmol, 545.71 μL, 8eq). The mixture was stirred at 50° C. for 2 h and concentrated to yieldN-[2-(5-fluoro-3-pyridyl)-8-isopropyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl]-1,2,3,4-tetrahydrocyclopenta[b]indol-2-amine(120 mg, 274.65 μmol, 70.1% yield, 97.8% purity) which was separated bychiral SFC (column: DAICEL CHIRALCEL OJ-H (250 mm*30 mm, 5 μm); mobilephase: [0.1% NH₃H₂O EtOH]; B %: 40%-40%) to yield peak 1 and peak 2.Peak 1 was concentrated under reduced pressure to yield a residue whichwas purified by preparative HPLC (column: Agela ASB 150*25 mm*5 um;mobile phase: [water (0.05% HCl)-ACN]; B %: 68%-98%, 8 min). The desiredfraction was lyophilized to yield an enantiomer (31.51 mg, 62.97 μmol,16.1% yield, 100.0% purity, 2HCl, SFC: R_(t)=5.092, ee=99.41%,[α]^(24.5) _(D)=+37.706 (MeOH, c=0.093 g/100 mL)) as a yellow solid. ¹HNMR (400 MHz, DMSO-d₆) δ ppm 10.94 (s, 1H), 9.43-9.40 (m, 2H), 8.71 (d,J=2.9 Hz, 1H), 8.45-8.41 (m, 1H), 8.15 (s, 1H), 7.35-7.31 (m, 2H),7.00-6.95 (m, 2H), 5.65-5.60 (m, 1H), 3.43-3.34 (m, 2H), 3.24-3.19 (m,2H), 3.03 (dd, J=6.6, 13.9 Hz, 1H), 1.37 (d, J=6.8 Hz, 6H); ES-LCMS m/z428.2 [M+H]⁺. Peak 2 was concentrated under reduced pressure to yield aresidue which was purified by preparative HPLC (column: Agela ASB 150*25mm*5 um; mobile phase: [water (0.05% HCl)-ACN]; B %: 68%-98%, 8 min).The desired fraction was lyophilized to yield the other enantiomer(24.54 mg, 49.04 μmol, 12.5% yield, 100.0% purity, 2HCl, SFC:R_(t)=6.749, ee=99.76%, [α]^(24.6) _(D)=−41.018 (MeOH, c=0.087 g/100mL)) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.93 (s, 1H),9.43-9.40 (m, 2H), 8.71 (d, J=2.7 Hz, 1H), 8.45-8.41 (m, 1H), 8.15 (s,1H), 7.35-7.31 (m, 2H), 7.02-6.95 (m, J=7.0, 15.1 Hz, 2H), 5.67-5.60 (m,1H), 3.41-3.34 (m, 2H), 3.24-3.21 (m, J=7.2, 14.1 Hz, 2H), 3.04-2.99(dd, J=6.4, 13.9 Hz, 1H), 1.37 (d, J=6.8 Hz, 6H); ES-LCMS m/z 428.2[M+H]⁺.

Example 200

Synthesis of I-264

Synthetic Scheme:

Step 1: tert-Butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

To a solution of(3R)—N-(8-isopropyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(300 mg, 726.08 μmol, 1 eq) in 1,4-dioxane (30 mL) was added DMAP(266.12 mg, 2.18 mmol, 3 eq) and Boc₂O (950.78 mg, 4.36 mmol, 1.00 mL, 6eq). The mixture was stirred at 110° C. for 10 h. To the mixture wasadded Boc₂O (950.78 mg, 4.36 mmol, 1.00 mL, 6 eq). The mixture wasstirred at 110° C. for another 16 h. The mixture was concentrated. Thecrude material was purified on silica gel column chromatography (frompure PE to PE/EtOAc=3/1, TLC: PE/EtOAc=5/1, R_(f)=0.7) to yieldtert-butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(420 mg, 637.71 μmol, 87.8% yield, 90.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.10-8.04 (m, 1H), 7.93 (s, 1H), 7.33 (d,J=7.3 Hz, 1H), 7.23-7.13 (m, 2H), 4.76-4.74 (m, 1H), 3.33-2.97 (m, 5H),2.59-2.51 (m, 3H), 2.37-2.27 (m, 1H), 2.24-2.26 (m, 1H), 1.63 (s, 9H),1.54 (s, 9H), 1.36 (d, J=6.8 Hz, 6H); ES-LCMS m/z 593.3 [M+H]⁺.

Step 2: tert-Butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropyl-2-methylsulfinyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

To a solution of tert-butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(400 mg, 607.34 μmol, 1 eq) in MeOH (20 mL) was added oxone (373.37 mg,607.34 μmol, 1 eq). The mixture was stirred at 15° C. for 3 h. Afterbeing filtered, to the filtration was added EtOAc (80 mL) and quenchedby addition with saturated Na₂SO₃ solution (50 mL). The aqueous layerwas extracted with EtOAc (30 mL×3) and the combined organic layers werewashed with brine (50 mL), dried over Na₂SO₄, filtered and concentrated.The crude material was purified on silica gel column chromatography(from PE/EtOAc=10/1 to 2/1, TLC: PE/EtOAc=5/1, R_(f)=0.2) to yieldtert-butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropyl-2-methylsulfinyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(250 mg, 403.70 μmol, 66.5% yield, 98.3% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.16 (s, 1H), 8.07 (d, J=8.1 Hz, 1H), 7.32(d, J=7.6 Hz, 1H), 7.24-7.14 (m, 2H), 4.85-4.83 (m, 1H), 3.40 (td,J=6.8, 13.8 Hz, 1H), 3.34-3.01 (m, 4H), 2.94 (d, J=2.7 Hz, 3H), 2.43 (d,J=12.0 Hz, 1H), 2.27 (d, J=8.1 Hz, 1H), 1.64 (s, 9H), 1.45-1.29 (m,15H); ES-LCMS m/z 631.3 [M+Na]⁺.

Step 3: tert-Butyl(3R)-3-[[8-isopropyl-2-(1-piperidyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate

A solution oftert-butyl(3R)-3-[tert-butoxycarbonyl-(8-isopropyl-2-methylsulfinyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(100 mg, 161.48 μmol, 1 eq) and piperidine (172.44 mg, 2.03 mmol, 0.2mL, 12.54 eq) in t-BuOH (1 mL) was irradiated and stirred at 120° C. for2 h under microwave. The mixture was concentrated to yield tert-butyl(3R)-3-[[8-isopropyl-2-(1-piperidyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(80 mg, 137.44 μmol, 85.1% yield, 91.0% purity) as a yellow solid. ¹HNMR (400 MHz, CDCl₃) δ ppm 8.11 (d, J=8.3 Hz, 1H), 7.57 (s, 1H), 7.35(d, J=7.1 Hz, 1H), 7.27 (br s, 1H), 7.23-7.17 (m, 1H), 6.26 (d, J=7.8Hz, 1H), 4.60-4.59 (m, 1H), 3.79 (d, J=4.6 Hz, 4H), 3.35 (d, J=5.1 Hz,3H), 3.27-3.14 (m, 3H), 3.02 (td, J=6.8, 13.8 Hz, 1H), 2.72 (dd, J=7.7,14.6 Hz, 1H), 2.25 (br s, 1H), 2.12 (d, J=8.3 Hz, 1H), 1.54-1.39 (m,12H), 1.28 (d, J=6.8 Hz, 6H); ES-LCMS m/z 530.4 [M+H]⁺.

Step 4:(3R)—N-[8-Isopropyl-2-(1-piperidyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-264)

A solution of tert-butyl(3R)-3-[[8-isopropyl-2-(1-piperidyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]amino]-1,2,3,4-tetrahydrocarbazole-9-carboxylate(80 mg, 137.44 μmol, 1 eq) and TFA (1.12 g, 9.83 mmol, 728.00 μL, 71.54eq) in DCM (5 mL) was stirred at 20° C. for 2 h. The mixture wasconcentrated. The residue was purified by preparative HPLC (column:Agela ASB 150*25 mm*5 μm; mobile phase: [water (0.05% HCl)-ACN]; B %:50%-80%, 8 min), followed by lyophilization to yield(3R)—N-[8-isopropyl-2-(1-piperidyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(34.59 mg, 71.55 μmol, 52.1% yield, 96.4% purity, HCl; Optical rotation:[α]^(22.2) _(D)=+82.409, (MeOH, c=0.096 g/100 mL)) as a yellow solid. ¹HNMR (400 MHz, CD₃OD) δ ppm 7.89 (s, 1H), 7.37 (d, J=7.8 Hz, 1H), 7.28(d, J=8.0 Hz, 1H), 7.04 (t, J=7.5 Hz, 1H), 7.00-6.93 (m, 1H), 4.69-4.57(m, 1H), 3.87 (br s, 4H), 3.28-3.16 (m, 2H), 2.97 (br s, 2H), 2.93-2.82(m, 1H), 2.37-2.15 (m, 2H), 1.74 (br s, 6H), 1.31 (d, J=6.8 Hz, 6H);ES-LCMS m/z 430.3 [M+H]⁺.

Example 201

Synthesis of I-265

Synthetic Scheme:

Step 1:2-(5-Fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carboxamide(I-265)

A solution of2-(5-fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carbonitrile(170 mg, 396.53 μmol, 1 eq) in HCl/MeOH (4 M in MeOH, 30 mL) was stirredat 80° C. for 12 h. The reaction mixture was concentrated under reducedpressure to yield a residue which was quenched by addition of saturatedaqueous NaHCO₃ (50 mL), extracted with EtOAc (30 mL×3). The combinedorganic layers were washed with brine (10 mL), dried over Na₂SO₄,filtered and concentrated under reduced pressure to yield a residue. Theresidue was purified by flash silica gel chromatography (fromPE/EtOAc=1/0 to 0/1, TLC: PE/EtOAc=1/1, R_(f)=0.11) to yield2-(5-fluoro-3-pyridyl)-4-[[(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-yl]amino]pyrazolo[1,5-a][1,3,5]triazine-8-carboxamide(60 mg, 135.61 μmol, 34.2% yield, 100% purity, [α]^(23.7) _(D)=+21.368(MeOH, c=0.088 g/100 mL)) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δppm 10.79 (s, 1H), 9.56-9.41 (m, 2H), 8.78-8.67 (m, 2H), 8.50 (s, 1H),7.52 (s, 1H), 7.45 (s, 1H), 7.34 (d, J=7.8 Hz, 1H), 7.28 (d, J=7.8 Hz,1H), 7.05-6.98 (m, 1H), 6.97-6.89 (m, 1H), 4.86 (s, 1H), 3.23-2.77 (m,4H), 2.22 (s, 2H); ES-LCMS m/z 443.1 [M+H]⁺.

Example 202

Synthesis of I-267

Synthetic Scheme:

Step 1: EthylN-[[4-(trifluoromethyl)-1H-pyrazol-5-yl]carbamothioyl]carbamate

A solution of 4-(trifluoromethyl)-1H-pyrazol-5-amine; hydrochloride (1g, 4.46 mmol, 1 eq, HCl) and ethyl N-(thioxomethylene)carbamate (585.47mg, 4.46 mmol, 1 eq) in DCM (10 mL) and DMF (10 mL) was stirred at 20°C. for 2.5 h. The reaction mixture was concentrated under reducedpressure to yield a residue of ethylN-[[4-(trifluoromethyl)-1H-pyrazol-5-yl]carbamothioyl]carbamate (1.1 g,3.90 mmol, 87.3% yield, crude purity) as a yellow solid which was usedin the next step without further purification. ¹H NMR (400 MHz, DMSO-d₆)δ ppm 11.52 (s, 1H), 11.06 (s, 1H), 8.34 (s, 1H), 4.21 (q, J=7.0 Hz,2H), 1.25 (t, J=7.0 Hz, 3H); ES-LCMS m/z 283.0 [M+H]⁺.

Step 2:2-Thioxo-8-(trifluoromethyl)-1H-pyrazolo[1,5-a][1,3,5]triazin-4-one

To a solution of ethylN-[[4-(trifluoromethyl)-1H-pyrazol-5-yl]carbamothioyl]carbamate (1.1 g,3.90 mmol, 1 eq) in CH₃CN (5 mL) was added K₂CO₃ (1.62 g, 11.69 mmol,3.0 eq). The mixture was stirred at 60° C. for 3 h. The reaction mixturewas quenched by AcOH solution (6 mL), diluted with H₂O (50 mL) andextracted with EtOAc (50 mL×3). The combined organic layers were driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto yield a residue. The residue was diluted with EtOAc/PE (2 mL/10 mL),filtered and collected the solid to yield2-thioxo-8-(trifluoromethyl)-1H-pyrazolo[1,5-a][1,3,5]triazin-4-one (900mg, 3.33 mmol, 85.4% yield, 87.3% purity) as a yellow solid which wasused in the next step without further purification. ¹H NMR (400 MHz,DMSO-d₆) δ ppm 12.65 (br s, 1H), 8.16 (s, 1H); ES-LCMS m/z 237.0 [M+H]⁺.

Step 3:2-Methylsulfanyl-8-(trifluoromethyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

To a solution of2-thioxo-8-(trifluoromethyl)-1H-pyrazolo[1,5-a][1,3,5]triazin-4-one (900mg, 3.33 mmol, 1 eq) in EtOH (15 mL) was added a solution of NaOH(266.12 mg, 6.65 mmol, 2 eq) in H₂O (6 mL) and then MeI (472.20 mg, 3.33mmol, 207.11 μL, 1 eq) was added dropwise. The mixture was stirred at20° C. for 1 h. The reaction mixture was acidified with 1N HCl solution(10 mL) and concentrated under reduced pressure to remove EtOH, solidwas formed, filtered and collected the solid to yield2-methylsulfanyl-8-(trifluoromethyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one(800 mg, 3.00 mmol, 90.2% yield, 93.8% purity) as a yellow solid whichwas used in the next step without further purification. ¹H NMR (400 MHz,CD₃OD) δ ppm 8.16 (s, 1H), 2.64 (s, 3H); ES-LCMS m/z 251.1 [M+H]⁺.

Step 4:4-Chloro-2-methylsulfanyl-8-(trifluoromethyl)pyrazolo[1,5-a][1,3,5]triazine

A solution of2-methylsulfanyl-8-(trifluoromethyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one(200 mg, 749.80 μmol, 1 eq) and DIEA (290.72 mg, 2.25 mmol, 391.80 μL,3.0 eq) in POCl₃ (34.43 g, 224.55 mmol, 20.87 mL, 299.47 eq) was stirredat 130° C. for 12 h under N₂ atmosphere. The reaction mixture wasconcentrated under reduced pressure to yield4-chloro-2-methylsulfanyl-8-(trifluoromethyl)pyrazolo[1,5-a][1,3,5]triazine(200 mg, 744.47 μmol, 99.3% yield, crude purity) as a black brown oilwhich was used in the next step without further purification. ¹H NMR(400 MHz, CDCl₃) δ ppm 8.29 (s, 1H), 2.66-2.64 (m, 3H).

Step 5:(3R)—N-[2-Methylsulfanyl-8-(trifluoromethyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine

To a solution of4-chloro-2-methylsulfanyl-8-(trifluoromethyl)pyrazolo[1,5-a][1,3,5]triazine(200 mg, 744.47 μmol, 1 eq) and(3R)-2,3,4,9-tetrahydro-1H-carbazol-3-amine (166.39 mg, 893.37 μmol, 1.2eq) in ACN (10 mL) was added DIEA (962.18 mg, 7.44 mmol, 1.30 mL, 10eq). The mixture was stirred at 60° C. for 3 h. The reaction mixture wasconcentrated under reduced pressure to yield a residue which waspurified on silica gel column chromatography (from PE/EtOAc=1/0 to 10/3,TLC: PE/EtOAc=3/1, R_(f)=0.32) to yield the product of(3R)—N-[2-methylsulfanyl-8-(trifluoromethyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(150 mg, 353.10 μmol, 47.4% yield, 98.5% purity) as a brown oil. ¹H NMR(400 MHz, CDCl₃) δ ppm 7.99 (s, 1H), 7.84 (br s, 1H), 7.47 (d, J=7.5 Hz,1H), 7.34 (d, J=7.8 Hz, 1H), 7.24-7.08 (m, 2H), 6.65 (d, J=9.3 Hz, 1H),4.82 (m, 1H), 3.31 (dd, J=5.4, 15.4 Hz, 1H), 3.03-2.84 (m, 3H), 2.60 (s,3H), 2.31-2.21 (m, 2H); ES-LCMS m/z 419.1 [M+H]⁺.

Step 6:(3R)—N-[2-(5-Fluoro-3-pyridyl)-8-(trifluoromethyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-267)

To a solution of(3R)—N-[2-methylsulfanyl-8-(trifluoromethyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(100 mg, 235.40 μmol, 1 eq) and (5-fluoro-3-pyridyl)boronic acid (99.51mg, 706.20 μmol, 3.0 eq) in 1,4-dioxane (6 mL) was added Pd(PPh₃)₄(27.20 mg, 23.54 μmol, 0.1 eq) and thiophene-2-carbonyloxycopper (89.78mg, 470.80 μmol, 2.0 eq). The mixture was purged with N₂ for 1 min andstirred at 120° C. for 5 h under microwave. The reaction mixture wasfiltered and concentrated under reduced pressure to yield a residuewhich was purified on silica gel column chromatography (fromPE/EtOAc=1/0 to 10/3, TLC: PE/EtOAc=3/1, R_(f)=0.38) and preparativeHPLC (column: DuraShell 150*25 mm*5 um; mobile phase: [water (0.05%HCl)-ACN]; B %: 45%-75%, 8 min), followed by lyophilization to yield theproduct of(3R)—N-[2-(5-fluoro-3-pyridyl)-8-(trifluoromethyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl]-2,3,4,9-tetrahydro-1H-carbazol-3-amine(36.04 mg, 66.30 μmol, 28.2% yield, 99.4% purity, 2HCl, Opticalrotation: ([α]^(24.3) _(D)=+51.474, (MeOH, c=0.048 g/100 mL)) as ayellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 9.48 (br s, 1H), 8.71 (d,J=8.8 Hz, 2H), 8.35 (s, 1H), 7.36 (d, J=7.8 Hz, 1H), 7.26 (d, J=8.3 Hz,1H), 7.06-6.99 (m, 1H), 6.97-6.88 (m, 1H), 4.91 (d, J=3.8 Hz, 1H), 3.29(d, J=5.5 Hz, 1H), 3.10-2.90 (m, 3H), 2.45-2.35 (m, 1H), 2.33-2.21 (m,1H); ES-LCMS m/z 468.2 [M+H]⁺.

Example 203

Synthesis of I-270

Synthetic Scheme:

Step 1: 5-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine

A mixture of 5-bromopyrimidine (1 g, 6.29 mmol, 1 eq),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(1.76 g, 6.93 mmol, 1.10 eq), Pd(dppf)Cl₂.CH₂Cl₂ (513.66 mg, 628.99μmol, 0.1 eq), KOAc (1.85 g, 18.87 mmol, 3 eq) in 1,4-dioxane (40 mL)and DMF (10 mL) was stirred at 100° C. for 2 h under N₂ atmosphere. Themixture was concentrated. The residue was purified on silica gel columnchromatography (from PE/EtOAc=1/0 to 3/1, TLC: PE/EtOAc=3/1, R_(f)=0.23)to yield 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (1.5g, 5.82 mmol, 92.6% yield, 80% purity) as red oil. ¹H NMR (400 MHz,CDCl₃) δ ppm 9.25 (s, 1H), 9.00 (br s, 2H), 1.33 (s, 12H).

Step 2:(3R)—N-(8-Isopropyl-2-pyrimidin-5-yl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(I-270)

A mixture of(3R)—N-(8-isopropyl-2-methylsulfanyl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(150 mg, 351.57 μmol, 1 eq),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (271.66 mg,1.05 mmol, 3 eq), Pd₂(dba)₃ (32.19 mg, 35.16 μmol, 0.1 eq) and copper(I) thiophene-2-carbonyloxy (134.08 mg, 703.15 μmol, 2 eq) in1,4-dioxane (4 mL) was stirred at 110° C. for 12 h under N₂ atmosphere.The mixture was concentrated to yield crude material which was purifiedon silica gel column chromatography (from PE/EtOAc=1/0 to 2/1, TLC:PE/EtOAc=3/1, R_(f)=0.25) to yield a gray solid which was slurry withMeCN/H₂O/DMF (3/1/0.5, 10 mL). After filtration, the filter cake waswashed with MeCN (10 mL×3) and dried under vacuum to yield(3R)—N-(8-isopropyl-2-pyrimidin-5-yl-pyrazolo[1,5-a][1,3,5]triazin-4-yl)-2,3,4,9-tetrahydro-1H-carbazol-3-amine(42.34 mg, 99.74 μmol, 28.4% yield, 100% purity, [α]^(24.5) _(D)=+83.175(DMSO, c=0.102 g/100 mL)) as a gray solid. ¹H NMR (400 MHz, DMSO) δ ppm10.75 (s, 1H), 9.57 (s, 2H), 9.26 (s, 1H), 9.01 (br s, 1H), 8.12 (s,1H), 7.31 (d, J=7.6 Hz, 1H), 7.24 (d, J=8.1 Hz, 1H), 7.01-6.94 (m, 1H),6.93-6.86 (m, 1H), 4.80-4.77 (m, 1H), 3.11-3.00 (m, 2H), 2.99-2.81 (m,3H), 2.18 (m, 2H), 1.34 (d, J=6.8 Hz, 6H); ES-LCMS m/z 425.2 [M+H]⁺.

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments that utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments that have been represented by way of example.

We claim:
 1. A method for treating a cancer in a patient in needthereof, wherein the cancer is selected from the group consisting ofbladder cancer, melanoma, ovarian cancer, head and neck squamous cellcarcinoma (HNSCC), lung cancer, kidney cancer, stomach cancer,esophageal cancer, and acute myeloid leukemia (AML), comprisingadministering to said patient a compound selected from:

or a pharmaceutically acceptable salt thereof.
 2. The method accordingto claim 1, wherein the compound is selected from

or a pharmaceutically acceptable salt thereof.
 3. The method accordingto claim 2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 4. The method accordingto claim 2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 5. The method accordingto claim 2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 6. The method accordingto claim 2, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 7. The method accordingto claim 1, wherein the compound, or a pharmaceutically acceptable saltthereof, is administered in combination with an anti-cancer, cytotoxin,or chemotherapeutic agent.
 8. The method according to claim 1, whereinthe compound, or a pharmaceutically acceptable salt thereof, isadministered in combination with an immuno-oncology agent.
 9. The methodaccording to claim 8, wherein the immuno-oncology agent is a PD-1antagonist.
 10. The method according to claim 8, wherein the compound,or a pharmaceutically acceptable salt thereof, is administered prior to,or concurrently with, or after, administration of the immuno-oncologyagent.
 11. A method for treating a bladder cancer in a patient in needthereof, comprising administering to said patient a compound selectedfrom:

or a pharmaceutically acceptable salt thereof.
 12. The method accordingto claim 11, wherein the compound is selected from

or a pharmaceutically acceptable salt thereof.
 13. The method accordingto claim 11, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 14. The method accordingto claim 11, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 15. The method accordingto claim 11, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 16. The method accordingto claim 11, wherein the compound is

or a pharmaceutically acceptable salt thereof.
 17. The method accordingto claim 11, wherein the compound, or a pharmaceutically acceptable saltthereof, is administered in combination with an anti-cancer, cytotoxin,or chemotherapeutic agent.
 18. The method according to claim 11, whereinthe compound, or a pharmaceutically acceptable salt thereof, isadministered in combination with an immuno-oncology agent.
 19. Themethod according to claim 18, wherein the immuno-oncology agent is aPD-1 antagonist.
 20. The method according to claim 18, wherein thecompound, or a pharmaceutically acceptable salt thereof, is administeredprior to, or concurrently with, or after, administration of theimmuno-oncology agent.